WO2019141250A1 - Pyridone-pyrimidine derivative acting as krasg12c mutein inhibitor - Google Patents

Abstract

Provided are a class of KRAS G12C mutein inhibitors, which relate in particular to a compound represented by formula (I), an isomer thereof, and a pharmaceutically acceptable salt thereof.

Description

Pyridone-pyrimidine derivatives as KRASG12C mutein inhibitors

This application claims the following priority:

CN201810055396.8, the application date is January 19, 2018;

CN201810712103.9, the application date is June 29, 2018.

Technical field

The present invention relates to a novel substituted pyridone pyrimidine derivative, in particular to a compound of the formula (I) or an isomer thereof, a pharmaceutically acceptable salt, and a compound of the formula (I) or an isomer thereof The use of a pharmaceutically acceptable salt and a pharmaceutical composition for the preparation of a medicament for the treatment of cancer.

Background technique

The first RAS oncogene was found in rat sarcoma, hence the name. The RAS protein is a product expressed by the RAS gene and refers to a closely related monomeric globulin consisting of 189 amino acids with a molecular weight of 21 kDa. The RAS protein can bind to guanine trinucleotide phosphate (GTP) or guanine dinucleotide phosphate (GDP). The active state of RAS protein has an effect on cell growth, differentiation, cytoskeleton, protein transport and secretion. Its activity is regulated by binding to GTP or GDP: when RAS protein binds to GDP, it is dormant, that is, "inactivated" state; when upstream specific cell growth factor is stimulated, RAS protein is induced Exchange GDP, combined with GTP, is called the "activation" state. The RAS protein that binds to GTP activates downstream proteins for signal transmission. The RAS protein itself has weak hydrolyzed GTP hydrolyzing activity and is capable of hydrolyzing GTP to GDP. This allows conversion from an activated state to an inactivated state. In this hydrolysis process, GAP (GTPase activating proteins) is also involved. It works with RAS proteins and greatly enhances its ability to hydrolyze GTP to GDP. Mutations in the RAS protein will affect its interaction with GAP, which affects its ability to hydrolyze GTP to GDP, making it active. The activated RAS protein continues to give downstream protein growth signals, ultimately leading to constant cell growth and differentiation, ultimately resulting in tumors. There are many members of the RAS gene family. Among them, the subfamilies closely related to various cancers include Kirsten rat sarcoma virus oncogene homolog (KRAS), Harvey rat sarcoma virus oncogenic homolog (HRAS) and nerve. Maternal tumor rat sarcoma virus oncogene homolog (NRAS). It has been found that approximately 30% of human tumors carry certain mutated RAS genes, with KRAS mutations being the most prominent, accounting for 86% of all RAS mutations. For KRAS mutations, the most common mutations occurred on residues 12 (G12), 13 glycine (G13) and 61 glutamine (Q61), with G12 mutations accounting for 83%.

The G12C mutation is a relatively common subtype of the KRAS gene mutation, which means that the 12th glycine is mutated to cysteine. KRAS G12C mutations are most common in lung cancer. According to data reported in the literature (Nat Rev Drug Discov 2014; 13: 828-851), KRAS G12C mutations account for about 10% of all lung cancer patients.

KRAS G12C mutant protein as a leading target, the current research is not a lot. The literature (Nature. 2013; 503: 548-551) reports a class of covalent binding inhibitors that target the KRAS G12C mutation, but such compounds are not enzymatically active and exhibit no activity at the cellular level. A class of compounds reported in the literature (Science 2016; 351: 604-608, Cancer Discov 2016; 6: 316-29) showed cell anti-proliferative activity at the muM level at the cellular level, but its metabolic stability was poor and its activity was difficult. Make further improvements. In recent years, Araxes Pharma has applied for several patents for KRAS G12C inhibitors. For example, WO2016164675 and WO2016168540 report that a class of quinazoline derivatives have high enzyme binding activity and exhibit cell proliferation resistance at the μM level. Its structure is stable and has certain selectivity. Amgen (WO2018119183A2) and AstraZeneca (WO2018206539) have published patents on KRAS G12C inhibitors in 2018, respectively, and Amgen's KRAS G12C inhibitor AMG 510 initiated a phase I clinical study in July 2018. Looking at the KRAS G12C inhibitors reported in the literature, they all have a fragment of acrylamide which acts as a cysteine residue on the Michael addition receptor and the KRASG12C mutein to form a covalently bound complex. In 2018, LiuYi et al., Cell (Matthew R. Janes, Yi Liu et al., Cell, 2018, 172, 578-589.) publicly reported a covalent binding inhibitor, ARS-1620, targeting the KRAS G12C mutation. It has good metabolic stability, exhibits nM-level cell anti-proliferative activity at the cellular level, and can effectively inhibit tumor growth in a pancreatic cancer MIA-Paca2 cell subcutaneous xenograft tumor model.

Compound Ex3 is disclosed in WO 2018/064510 A1, but no characterization data and test results are given.

Summary of the invention

The present invention provides a compound of the formula (I), a pharmaceutically acceptable salt thereof or an isomer thereof,

among them,

Ring A is selected from 3 to 8 membered heterocycloalkyl, and said 3 to 8 membered heterocycloalkyl is optionally substituted by 1, 2 or 3 R;

R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from the group consisting of H, halogen, OH, NH ₂ , CN, C _1-6 alkyl and C _1-6 heteroalkyl, said C _{1- 6} alkyl and C _1-6 heteroalkyl are optionally substituted by 1, 2 or 3 R;

Or, R ₁ and R _{2 are} joined together to form a ring B;

Or R ₂ and R _{3 are} joined together to form a ring B;

Or R ₃ and R _{4 are} joined together to form a ring B;

Or R ₄ and R _{5 are} joined together to form a ring B;

Ring B is selected from the group consisting of phenyl, C _5-6 cycloalkenyl, 5- to 6-membered heterocycloalkenyl, and 5- to 6-membered heteroaryl, phenyl, C _5-6 cycloalkenyl, and 5 to 6-membered hetero a cycloalkenyl group, a 5- to 6-membered heteroaryl group, optionally substituted by 1, 2 or 3 R _a ;

R _{a is} selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-6 alkyl and C _1-6 heteroalkyl, the C _1-6 alkyl and C _1-6 heteroalkyl optionally being 1, 2 Or 3 R substitutions;

R ₆ is selected from H, halogen and C _1-6 alkyl, said C _1-6 alkyl optionally substituted with 1, 2 or 3 R <

R _{7 is} selected from the group consisting of H, CN, NH ₂ , C _1-8 alkyl, C _1-8 heteroalkyl, 4-6-membered heterocycloalkyl, 5- to 6-membered heteroaryl, and C _5-6 cycloalkyl , the C _1-8 alkyl group, the C _1-8 heteroalkyl group, the 4-6 membered heterocycloalkyl group, the 5- to 6-membered heteroaryl group, and the C _5-6 cycloalkyl group are selected from the group consisting of 1, 2 Or 3 R substitutions;

L is selected from the group consisting of a single bond, -NH-, -S-, -O-, -C(=O)-, -C(=S)-, -CH ₂ -, -CH(R _b )-, and -C( R _b ) ₂ -;

L' is selected from the group consisting of a single bond and -NH-;

R _{b is} selected from C _1-3 alkyl and C _1-3 heteroalkyl, and the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted by 1, 2 or 3 R;

R _{8 is} selected from H, C _1-6 alkyl and C _1-6 heteroalkyl, and the C _1-6 alkyl and C _1-6 heteroalkyl are optionally substituted by 1, 2 or 3 R;

R is selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-6 alkyl, C _1-6 heteroalkyl, and C _3-6 membered cycloalkyl, said C _1-6 alkyl, C _1-6 hetero An alkyl group and a C _3-6 membered cycloalkyl group are optionally substituted by 1, 2 or 3 R';

R' is selected from the group consisting of: F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , CH ₃ O, CF ₃ , CHF ₂ , CH ₂ F, cyclopropyl, propyl, iso Propyl, N(CH ₃ ) ₂ , NH(CH ₃ );

"Heter" means a hetero atom or a hetero atom group, the 3- to 8-membered heterocycloalkyl group, a C _1-6 heteroalkyl group, a 5- to 6-membered heterocycloalkenyl group, a 5- to 6-membered heteroaryl group, and C _{1 - 8} The "hetero" of a heteroalkyl group, a 4-6 membered heterocycloalkyl group, and a _C1-3 heteroalkyl group are each independently selected from -C(=O)N(R)-, -N(R)-, -NH. -, N, -O-, -S-, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O ₂ - and -N(R)C(=O)N(R)-;

In any of the above cases, the number of heteroatoms or heteroatoms is independently selected from 1, 2 and 3.

In some aspects of the invention, the above R is selected from the group consisting of F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , CH ₃ O, CF ₃ , CHF ₂ , CH ₂ F, ring Propyl, propyl, isopropyl, N(CH ₃ ) ₂ , NH(CH ₃ ) and N(CH ₂ CH ₃ ) ₂ .

In some embodiments of the invention, ring A is selected from the group consisting of aziridine, azetidine, pyrrolidine, piperidinyl, piperazinyl, 1,4-diazacycloheptyl, and 3,6- Diazabicyclo[3.2.0]heptane, the aziridine, azetidine, pyrrolidine, piperidinyl, piperazinyl, 1,4-diazacycloheptyl and 3, 6-diazabicyclo[3.2.0]heptane is optionally substituted by 1, 2 or 3 R.

In some embodiments of the invention, the above R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH and CH ₃ NH(C=O)O, the CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH and CH ₃ NH(C=O)O are optionally substituted by 1, 2 or 3 R.

In some embodiments of the invention, the above R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ N(C=O)O and CH ₃ NH(C=O)O.

In some embodiments of the invention, the above ring B is selected from pyrazolyl, imidazolyl, pyrrolyl, thienyl, furyl, triazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, benzene a pyridyl group, a pyridalyl group, a pyridazinyl group Or oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, phenyl, pyridyl, pyrimidinyl, pyridazinyl, triazinyl, morpholinyl, cyclopentenyl and cyclohexenyl optionally Replaced by 1, 2 or 3 R _a .

In some aspects of the present invention, the above R _a is selected from F, Cl, Br, I, OH, NH 2, CN, CH 3, CH 3 CH 2, (CH 3) 2 CH, CH 3 O, CH 3 C (=O).

In some embodiments of the invention, Ring B is selected from the group consisting of phenyl, pyrazolyl, 1-methyl-1H-pyrazolyl, and 1-(1H-pyrazol-1-yl)ethanone.

In some embodiments of the invention, the above R _{6 is} selected from the group consisting of H, F, Cl, Br, I, and C _1-3 alkyl, and the C _1-3 alkyl group is optionally substituted with 1, 2 or 3 R.

In some embodiments of the invention, the above R _{6 is} selected from the group consisting of H, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ , CH ₂ F.

In some embodiments of the invention, R ₇ above is selected from the group consisting of H, CN, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, morpholinyl, piperidinyl, azetidinyl, Azacyclopentyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, cyclohexane, cyclopentyl, phenyl, pyridyl, pyridazinyl, Pyrimidinyl, pyrazinyl, said C _1-6 alkyl, C _1-6 heteroalkyl, morpholinyl, piperidinyl, azetidinyl, azetidinyl, pyrazolyl, Imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, cyclohexane, cyclopentyl, phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl are optionally 2 or 3 R substitutions.

所述

任选被1、2或3个R取代。 In some embodiments of the invention, the above R _{7 is} selected from the group consisting of H, CH ₃ , CN, NH ₂ ,

Said

Optionally substituted by 1, 2 or 3 R.

In some embodiments of the invention, the above R _{7 is} selected from the group consisting of H, CH ₃ , CN, NH ₂ ,

In some embodiments of the invention, the above R _{8 is} selected from the group consisting of H, C _1-4 alkyl and C _1-4 heteroalkyl, the C _1-4 alkyl and C _1-4 heteroalkyl optionally being 1 , 2 or 3 R substitutions.

In some embodiments of the invention, the above R _{8 is} selected from the group consisting of H, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CHCH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ NCH ₂ and CH ₃ NHCH ₂ .

选自

其中，R ₉选自H和C _1-3烷基。 In some aspects of the invention, the structural unit

Selected from

Wherein R _{9 is} selected from the group consisting of H and C _1-3 alkyl.

选自

In some aspects of the invention, the structural unit

Selected from

选自H、CN、CH ₃、CH ₃CH ₂、(CH ₃) ₂CH、(CH ₃) ₂N、(CH ₃) ₂NCH ₂、

In some aspects of the invention, the structural unit

Selected from H, CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ NCH ₂ ,

选自

In some aspects of the invention, the structural unit

Selected from

In some aspects of the invention, the above R is selected from the group consisting of F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , CH ₃ O, CF ₃ , CHF ₂ , CH ₂ F, ring Propyl, propyl, isopropyl, N(CH ₃ ) ₂ , NH(CH ₃ ) and N(CH ₂ CH ₃ ) ₂ , other variables are as defined in the present invention.

In some embodiments of the invention, ring A is selected from the group consisting of aziridine, azetidine, pyrrolidine, piperidinyl, piperazinyl, 1,4-diazacycloheptyl, and 3,6- Diazabicyclo[3.2.0]heptane, the aziridine, azetidine, pyrrolidine, piperidinyl, piperazinyl, 1,4-diazacycloheptyl and 3, 6-diazabicyclo[3.2.0]heptane is optionally substituted by 1, 2 or 3 R, other variables being as defined herein.

In some embodiments of the invention, the above R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH and CH ₃ NH(C=O)O, the CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH and CH ₃ NH(C=O)O are optionally substituted by 1, 2 or 3 R, and other variables are as defined in the present invention.

In some embodiments of the invention, the above R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ N(C=O)O and CH ₃ NH(C=O)O, other variables As defined by the present invention.

In some embodiments of the invention, the above ring B is selected from pyrazolyl, imidazolyl, pyrrolyl, thienyl, furyl, triazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, benzene a pyridyl group, a pyridalyl group, a pyridazinyl group Or oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, phenyl, pyridyl, pyrimidinyl, pyridazinyl, triazinyl, morpholinyl, cyclopentenyl and cyclohexenyl optionally It is substituted by 1, 2 or 3 R _a and the other variables are as defined in the present invention.

In some aspects of the present invention, the above R _a is selected from F, Cl, Br, I, OH, NH 2, CN, CH 3, CH 3 CH 2, (CH 3) 2 CH, CH 3 O, CH 3 C (=O), other variables are as defined by the present invention.

In some embodiments of the invention, the above ring B is selected from the group consisting of phenyl, pyrazolyl, 1-methyl-1H-pyrazolyl and 1-(1H-pyrazol-1-yl)ethanone, and other variables such as The invention is defined.

In some embodiments of the invention, the above R _{6 is} selected from the group consisting of H, F, Cl, Br, I, and C _1-3 alkyl, and the C _1-3 alkyl group is optionally substituted with 1, 2 or 3 R. Other variables are as defined by the present invention.

In some aspects of the invention, R ₆ above is selected from the group consisting of H, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ , CH ₂ F, and other variables are as defined herein.

In some embodiments of the invention, R ₇ above is selected from the group consisting of H, CN, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, morpholinyl, piperidinyl, azetidinyl, Azacyclopentyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, cyclohexane, cyclopentyl, phenyl, pyridyl, pyridazinyl, Pyrimidinyl, pyrazinyl, said C _1-6 alkyl, C _1-6 heteroalkyl, morpholinyl, piperidinyl, azetidinyl, azetidinyl, pyrazolyl, Imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, cyclohexane, cyclopentyl, phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl are optionally 2 or 3 R substitutions, other variables as defined by the present invention.

所述

任选被1、2或3个R取代，其他变量如本发明所定义。 In some embodiments of the invention, the above R _{7 is} selected from the group consisting of H, CH ₃ , CN, NH ₂ ,

Said

Optionally substituted by 1, 2 or 3 R, other variables are as defined by the present invention.

其他变量如本发明所定义。 In some embodiments of the invention, the above R _{7 is} selected from the group consisting of H, CH ₃ , CN, NH ₂ ,

Other variables are as defined by the present invention.

In some embodiments of the invention, the above R _{8 is} selected from the group consisting of H, C _1-4 alkyl and C _1-4 heteroalkyl, the C _1-4 alkyl and C _1-4 heteroalkyl optionally being 1 , 2 or 3 R substitutions, other variables as defined by the present invention.

In some embodiments of the invention, the above R _{8 is} selected from the group consisting of H, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CHCH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ NCH ₂ and CH ₃ NHCH ₂ , other variables are as defined in the present invention.

选自

其中，R ₉选自H和C _1-3烷基，其他变量如本发明所定义。 In some aspects of the invention, the structural unit

Selected from

Wherein R _{9 is} selected from the group consisting of H and C _1-3 alkyl, and other variables are as defined in the present invention.

选自

其他变量如本发明所定义。 In some aspects of the invention, the structural unit

Selected from

Other variables are as defined by the present invention.

选自H、CN、CH ₃、CH ₃CH ₂、(CH ₃) ₂CH、(CH ₃) ₂N、(CH ₃) ₂NCH ₂、

其他变量如本发明所定义。 In some aspects of the invention, the structural unit

Selected from H, CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ NCH ₂ ,

Other variables are as defined by the present invention.

选自

其他变量如本发明所定义。 In some aspects of the invention, the structural unit

Selected from

Other variables are as defined by the present invention.

In some embodiments of the invention, the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof, is selected from

Wherein L, R ₁ , R ₂ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ and R _{9 are} as defined in the present invention.

In some embodiments of the invention, the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof, is selected from

Wherein R ₁ , R ₂ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , L and ring B are as defined in the present invention.

In some embodiments of the invention, the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof, is selected from

Wherein R ₁ , R ₂ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , L, R ₉ and R _{a are} as defined in the present invention.

The present invention also provides a compound of the formula: a pharmaceutically acceptable salt thereof or an isomer thereof, selected from the group consisting of

In some embodiments of the invention, the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof, is selected from

The present invention also provides the use of the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof for the preparation of a medicament for treating cancer.

In some aspects of the invention, the cancer includes lung cancer, lymphoma, esophageal cancer, ovarian cancer, pancreatic cancer, rectal cancer, glioma, cervical cancer, urothelial carcinoma, gastric cancer, endometrial cancer, liver cancer. , cholangiocarcinoma, breast cancer, colon cancer, leukemia and melanoma.

Still other aspects of the invention are arbitrarily combined from the above variables.

Technical effect

The compound of the present invention comprises a substituted pyridone and pyrimidine derivative, which has high cell anti-proliferative activity against KRAS G12C mutein, and has a weak cell activity against wild type, showing good selectivity, showing that Classes of compounds have better safety as potential therapeutic agents. The mother nucleus of the compound of the present invention is a pyridone-pyrimidine structure, which has a relatively high polarity and a high solubility, and the substituent on the aromatic ring on the left side has a significant influence on the activity, selectivity and pharmacokinetic properties of the compound. This type of structure has high chemical stability and also exhibits high metabolic stability in vitro. In the rat pharmacokinetic evaluation experiment, the compounds of the present invention showed higher exposure and better oral availability than the reference compound ARS-1620. The compound of the present invention exhibited a more significant antitumor effect than the reference compound ARS-1620 in the human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model and the human pancreatic cancer x-MIA-PaCa2 subcutaneous xenograft model. In addition, the structure of pyridone and pyrimidine is reported in the literature, and it is difficult to carry out substitution or derivatization chemistry in its structure. The present invention also provides a novel pyridone-pyrimidine structure synthesis method which can synthesize a series of derivatives by first constructing a pyridone structure and then constructing a pyrimidine ring starting from different substituted amines. This method has not been reported in the literature and is an effective method for synthesizing such compounds.

Definition and description

Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered undefined or unclear without a particular definition, but should be understood in the ordinary sense. When a trade name appears in this document, it is intended to refer to its corresponding commodity or its active ingredient. The term "pharmaceutically acceptable" as used herein is intended to mean that those compounds, materials, compositions and/or dosage forms are within the scope of sound medical judgment and are suitable for use in contact with human and animal tissues. Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to a salt of a compound of the invention prepared from a compound having a particular substituent found in the present invention and a relatively non-toxic acid or base. When a relatively acidic functional group is contained in the compound of the present invention, a base addition salt can be obtained by contacting a neutral amount of such a compound with a sufficient amount of a base in a neat solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When a relatively basic functional group is contained in the compound of the present invention, an acid addition salt can be obtained by contacting a neutral form of such a compound with a sufficient amount of an acid in a neat solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogencarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and an organic acid salt, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine, etc.) And salts of organic acids such as glucuronic acid. Certain specific compounds of the invention contain both basic and acidic functional groups which can be converted to any base or acid addition salt.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing an acid group or a base by conventional chemical methods. In general, such salts are prepared by reacting these compounds in water or an organic solvent or a mixture of the two via a free acid or base form with a stoichiometric amount of a suitable base or acid.

In addition to the form of the salt, the compounds provided herein also exist in the form of prodrugs. Prodrugs of the compounds described herein are readily chemically altered under physiological conditions to convert to the compounds of the invention. Furthermore, prodrugs can be converted to the compounds of the invention by chemical or biochemical methods in an in vivo setting.

Certain compounds of the invention may exist in unsolvated or solvated forms, including hydrated forms. In general, the solvated forms are equivalent to the unsolvated forms and are included within the scope of the invention.

The compounds of the invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including the cis and trans isomers, the (-)- and (+)-p-enantiomers, the (R)- and (S)-enantiomers, and the diastereomeric a conformation, a (D)-isomer, a (L)-isomer, and a racemic mixture thereof, and other mixtures, such as enantiomerically or diastereomeric enriched mixtures, all of which belong to It is within the scope of the invention. Additional asymmetric carbon atoms may be present in the substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the invention.

Unless otherwise indicated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of one another.

Unless otherwise indicated, the term "cis-trans isomer" or "geometric isomer" is caused by the inability to freely rotate a single bond due to a double bond or a ring-forming carbon atom.

Unless otherwise indicated, the term "diastereomer" refers to a stereoisomer in which the molecule has two or more chiral centers and the molecules are in a non-mirrored relationship.

Unless otherwise indicated, "(D)" or "(+)" means dextrorotatory, "(L)" or "(-)" means left-handed, "(DL)" or "(±)" means racemic.

和楔形虚线键

表示一个立体中心的绝对构型，用直形实线键

和直形虚线键

表示立体中心的相对构型，用波浪线

表示楔形实线键

或楔形虚线键

或用波浪线

表示直形实线键

和直形虚线键

Wedge solid key unless otherwise stated

And wedge-shaped dashed keys

Represents the absolute configuration of a solid center with straight solid keys

And straight dashed keys

Indicates the relative configuration of the stereocenter, using wavy lines

Indicates a wedge solid key

Or wedge-shaped dotted key

Or with wavy lines

Represents a straight solid key

And straight dashed keys

The compounds of the invention may be present in particular. Unless otherwise indicated, the terms "tautomer" or "tautomeric form" mean that the different functional isomers are in dynamic equilibrium at room temperature and can be rapidly converted into each other. If tautomers are possible (as in solution), the chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also known as prototropic tautomers) include interconversions by proton transfer, such as keto-enol isomerization and imine-enes. Amine isomerization. The valence tautomer includes the mutual transformation of some of the bonding electrons. A specific example of keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise indicated, the terms "enriched in one isomer", "isomer enriched", "enriched in one enantiomer" or "enantiomeric enriched" refer to one of the isomers or pairs The content of the oligo is less than 100%, and the content of the isomer or enantiomer is 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, 98% or more, 99% or more, 99.5% or more, 99.6% or more, 99.7% or more, 99.8% or more, or greater than or equal to 99.9%.

Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the two isomers or the relative percentages of the two enantiomers. For example, if one of the isomers or enantiomers is present in an amount of 90% and the other isomer or enantiomer is present in an amount of 10%, the isomer or enantiomeric excess (ee value) is 80%. .

The optically active (R)- and (S)-isomers as well as the D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary wherein the resulting mixture of diastereomers is separated and the auxiliary group cleaved to provide pure The desired enantiomer. Alternatively, when a molecule contains a basic functional group (e.g., an amino group) or an acidic functional group (e.g., a carboxyl group), a diastereomeric salt is formed with a suitable optically active acid or base, followed by conventional methods well known in the art. The diastereomers are resolved and the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomers is generally accomplished by the use of chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (eg, formation of an amino group from an amine). Formate). The compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, radiolabeled compounds can be used, such as tritium ⁽³ H), iodine -125 ⁽¹²⁵ I) or ^C-14 (14 C). For another example, hydrogen can be replaced by heavy hydrogen to form a deuterated drug. The bond composed of barium and carbon is stronger than the bond composed of common hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have reduced side effects and increased drug stability. Enhance the efficacy and prolong the biological half-life of the drug. Alterations of all isotopic compositions of the compounds of the invention, whether radioactive or not, are included within the scope of the invention. "Optional" or "optionally" means that the subsequently described event or condition may, but is not necessarily, to occur, and that the description includes instances in which the event or condition occurs and instances in which the event or condition does not occur.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, and may include variants of heavy hydrogen and hydrogen, as long as the valence of the particular atom is normal and the substituted compound is stable. of. When the substituent is oxygen (ie, =0), it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on the aromatic group. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the kind and number of substituents may be arbitrary on the basis of chemically achievable.

When any variable (eg, R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 R, the group may optionally be substituted with at most two R, and each case has an independent option. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

When the number of one linking group is 0, such as -(CRR) ₀ -, it indicates that the linking group is a single bond.

When one of the variables is selected from a single bond, it means that the two groups to which it is attached are directly linked. For example, when L represents a single bond in A-L-Z, the structure is actually A-Z.

When a substituent is vacant, it means that the substituent is absent. For example, when X is vacant in A-X, the structure is actually A. When the listed substituents are not indicated by which atom is attached to the substituted group, such a substituent may be bonded through any atom thereof, for example, a pyridyl group as a substituent may be passed through any one of the pyridine rings. A carbon atom is attached to the substituted group.

中连接基团L为-M-W-，此时-M-W-既可以按与从左往右的读取顺序相同的方向连接环A和环B构成

也可以按照与从左往右的读取顺序相反的方向连接环A和环B构成

所述连接基团、取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。 When the listed linking group does not indicate its direction of attachment, its connection direction is arbitrary, for example,

The medium linking group L is -MW-, and at this time, -MW- can be connected in the same direction as the reading order from left to right to form ring A and ring B.

It is also possible to connect the ring A and the ring B in a direction opposite to the reading order from left to right.

Combinations of the linking groups, substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, the term "hetero" denotes a hetero atom or a hetero atomic group (ie, a radical containing a hetero atom), including atoms other than carbon (C) and hydrogen (H), and radicals containing such heteroatoms, including, for example, oxygen (O). ), nitrogen (N), sulfur (S), silicon (Si), germanium (Ge), aluminum (Al), boron (B), -O-, -S-, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O) ₂ -, and optionally substituted -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O) ₂ N(H)- or -S(=O)N(H)-.

Unless otherwise specified, "ring" means substituted or unsubstituted cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl or heteroaryl. The ring includes a single ring, and also includes a bicyclic or polycyclic ring system such as a spiro ring, a ring and a bridge ring. The number of atoms on the ring is usually defined as the number of elements of the ring. For example, "5 to 7-membered ring" means 5 to 7 atoms arranged in a circle. Unless otherwise specified, the ring optionally contains from 1 to 3 heteroatoms. Thus, the "5-7 membered ring" includes, for example, phenyl, pyridyl and piperidinyl; on the other hand, the term "5-7 membered heterocycloalkyl" includes pyridyl and piperidinyl, but does not include phenyl. The term "ring" also includes ring systems containing at least one ring, each of which "ring" independently conforms to the above definition.

Unless otherwise specified, the term "alkyl" is used to mean a straight or branched saturated hydrocarbon group, and in some embodiments, the alkyl group is a C _1-12 alkyl group; in other embodiments The alkyl group is a C _1-6 alkyl group; in other embodiments, the alkyl group is a C _1-3 alkyl group. It may be monosubstituted (such as -CH ₂ F) or polysubstituted (such as -CF ₃ ), and may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). . Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl) And t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl and the like.

Unless otherwise specified, "alkenyl" is used to indicate a straight or branched hydrocarbon group containing one or more carbon-carbon double bonds, and the carbon-carbon double bond may be located at any position of the group. In some embodiments, the alkenyl group is a C _2-8 alkenyl group; in other embodiments, the alkenyl group is a C _2-6 alkenyl group; in other embodiments, the alkenyl group is C _2-4 alkenyl. It may be monosubstituted or polysubstituted, and may be monovalent, divalent or multivalent. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl and the like.

Unless otherwise specified, the term "heteroalkyl", by itself or in conjunction with another term, denotes a stable straight or branched alkyl radical or a combination thereof consisting of a number of carbon atoms and at least one heteroatom or heteroatom. Things. In some embodiments, the heteroatoms are selected from the group consisting of B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatoms are optionally quaternized. In other embodiments, the heteroatom group is selected from the group consisting of -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O) ₂ -, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O) ₂ N(H)- and -S(=O)N( H)-. In some embodiments, the heteroalkyl group is a _C1-6 heteroalkyl group; in other embodiments, the heteroalkyl group is a _C1-3 heteroalkyl group. A heteroatom or heteroatom can be located at any internal position of a heteroalkyl group, including the position at which the alkyl group is attached to the rest of the molecule, but the terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkyl) Oxyl) is a conventional expression and refers to those alkyl groups which are attached to the remainder of the molecule through an oxygen atom, an amino group or a sulfur atom, respectively. Examples of heteroalkyl groups include, but are not limited to, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ CH ₃ , -OCH ₂ (CH ₃ ) ₂ , -CH ₂ -CH ₂ -O-CH ₃ , -NHCH ₃ , -N(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )(CH ₂ CH ₃ ), -CH ₂ -CH ₂ -NH-CH ₃ , -CH ₂ -CH ₂ -N ( CH ₃ )-CH ₃ , -SCH ₃ , -SCH ₂ CH ₃ , -SCH ₂ CH ₂ CH ₃ , -SCH ₂ (CH ₃ ) ₂ , -CH ₂ -S-CH ₂ -CH ₃ , -CH ₂ - CH ₂ , -S(=O)-CH ₃ , -CH ₂ -CH ₂ -S(=O) ₂ -CH ₃ , -CH=CH-O-CH ₃ , -CH ₂ -CH=N-OCH ₃ And -CH=CH-N(CH ₃ )-CH ₃ . Up to two heteroatoms may be consecutive, for example, -CH ₂ -NH-OCH _3.

Unless otherwise specified, the term "heteroalkenyl", by itself or in conjunction with another term, denotes a stable straight or branched alkenyl radical or a combination thereof consisting of a number of carbon atoms and at least one heteroatom or heteroatom. Things. In some embodiments, the heteroatoms are selected from the group consisting of B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatoms are optionally quaternized. In other embodiments, the heteroatom group is selected from the group consisting of -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O) ₂ -, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O) ₂ N(H)- and -S(=O)N( H)-. In some embodiments, the heteroalkenyl group is a C _2-6 heteroalkenyl group; in other embodiments, the heteroalkyl group is a C _2-4 heteroalkenyl group. The hetero atom or heteroatom group may be located at any internal position of the heteroalkenyl group, including the position at which the alkenyl group is attached to the rest of the molecule, but the terms "alkenyloxy", "alkenylamino" and "alkenylthio" are customary. By expression, it is meant those alkenyl groups which are attached to the remainder of the molecule through an oxygen atom, an amino group or a sulfur atom, respectively. Examples of heteroalkenyl groups include, but are not limited to, -O-CH=CH ₂ , -O-CH=CHCH ₃ , -O-CH=C(CH ₃ ) ₂ , -CH=CH-O-CH ₃ , -O- CH=CHCH ₂ CH ₃ , -CH ₂ -CH=CH-OCH ₃ , -NH-CH=CH ₂ , -N(CH=CH ₂ )-CH ₃ , -CH=CH-NH-CH ₃ , -CH =CH-N(CH ₃ ) ₂ , -S-CH=CH ₂ , -S-CH=CHCH ₃ , -S-CH=C(CH ₃ ) ₂ , -CH ₂ -S-CH=CH ₂ ,- S(=O)-CH=CH ₂ and -CH=CH-S(=O) ₂ -CH ₃ . Up to two heteroatoms may be consecutive, such as _{-CH = CH-NH-OCH 3} .

至多两个杂原子可以是连续的，例如

Unless otherwise specified, the term "heteroalkynyl", by itself or in conjunction with another term, denotes a stable straight or branched alkynyl radical or a combination thereof consisting of a number of carbon atoms and at least one heteroatom or heteroatom. Things. In some embodiments, the heteroatoms are selected from the group consisting of B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatoms are optionally quaternized. In other embodiments, the heteroatom group is selected from the group consisting of -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O) ₂ -, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O) ₂ N(H)- and -S(=O)N( H)-. In some embodiments, the heteroalkynyl group is a C _2-6 heteroalkynyl group; in other embodiments, the heteroalkyl group is a C _2-4 heteroalkynyl group. The hetero atom or heteroatom group may be located at any internal position of the heteroalkynyl group, including the position at which the alkynyl group is attached to the rest of the molecule, but the terms "alkynyloxy", "alkynylamino" and "alkynylthio" are customary. By expression, it is meant those alkynyl groups attached to the remainder of the molecule through an oxygen atom, an amino group or a sulfur atom, respectively. Examples of heteroalkynyl groups include, but are not limited to,

Up to two heteroatoms can be continuous, for example

Unless otherwise specified, "cycloalkyl" includes any stable cyclic alkyl group including monocyclic, bicyclic or tricyclic systems wherein the bicyclic and tricyclic systems include spiro, co and ring. In some embodiments, the cycloalkyl group is a C _3-8 cycloalkyl group; in other embodiments, the cycloalkyl group is a C _3-6 cycloalkyl group; in other embodiments, the The cycloalkyl group is a C _5-6 cycloalkyl group. It may be monosubstituted or polysubstituted, and may be monovalent, divalent or multivalent. Examples of such cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2]bicyclooctane, [4.4.0] Dicyclodecane and the like.

Unless otherwise specified, "cycloalkenyl" includes any stable cyclic alkenyl group containing one or more unsaturated carbon-carbon double bonds at any position of the group, including monocyclic, bicyclic or tricyclic The system wherein the bicyclic and tricyclic systems include spiro, parallel and bridged rings, but any ring of this system is non-aromatic. In some embodiments, the cycloalkenyl group is a C _3-8 cycloalkenyl group; in other embodiments, the cycloalkenyl group is a C _3-6 cycloalkenyl group; in other embodiments, the The cycloalkenyl group is a C _5-6 cycloalkenyl group. It may be monosubstituted or polysubstituted, and may be monovalent, divalent or multivalent. Examples of such cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclohexenyl, and the like.

Unless otherwise specified, "cycloalkynyl" includes any stable cyclic alkynyl group containing one or more carbon-carbon triple bonds at any position of the group, including monocyclic, bicyclic or tricyclic systems, wherein Bicyclic and tricyclic systems include spiro, parallel and bridging rings. It may be monosubstituted or polysubstituted, and may be monovalent, divalent or multivalent.

Unless otherwise specified, the term "heterocycloalkyl", by itself or in conjunction with other terms, denotes a cyclized "heteroalkyl", respectively, which includes monocyclic, bicyclic, and tricyclic systems, wherein the bicyclic and tricyclic systems include spiro rings, And ring and bridge ring. Further, in the case of the "heterocycloalkyl group", a hetero atom may occupy a position where a heterocycloalkyl group is bonded to the rest of the molecule. In some embodiments, the heterocycloalkyl group is a 4-6 membered heterocycloalkyl group; in other embodiments, the heterocycloalkyl group is a 5-6 membered heterocycloalkyl group. Examples of heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thioheterobutyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophene) -2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxoalkyl , dithiaalkyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl or oxygen Heterocyclic heptane.

Unless otherwise specified, the term "heterocyclenyl", by itself or in conjunction with other terms, denotes a cyclized "heteroalkenyl", respectively, which includes monocyclic, bicyclic, and tricyclic systems, wherein the bicyclic and tricyclic systems include spiro rings, Rings and bridge rings, but any ring of this system is non-aromatic. Furthermore, in the case of the "heterocyclenyl", a heteroatom can occupy the position of attachment of the heterocyclenyl group to the rest of the molecule. In some embodiments, the heterocycloalkenyl is 4 to 6 membered heterocycloalkenyl; in other embodiments, the heterocycloalkenyl is 5 to 6 membered heterocycloalkenyl. Examples of heterocycloalkenyl groups include, but are not limited to,

Unless otherwise specified, the term "heterocycloalkynyl", by itself or in conjunction with other terms, denotes a cyclized "heteroalkynyl" group, respectively, which includes monocyclic, bicyclic, and tricyclic systems, wherein the bicyclic and tricyclic systems include spiro rings, And ring and bridge ring. Further, in the case of the "heterocycloalkynyl group", a hetero atom may occupy a position where a heterocyclic alkynyl group is bonded to the rest of the molecule. In some embodiments, the heterocycloalkynyl group is a 4 to 6 membered heterocycloalkynyl group; in other embodiments, the heterocycloalkynyl group is a 5 to 6 membered heterocycloalkynyl group. Unless otherwise specified, the term "halo" or "halogen", by itself or as part of another substituent, denotes a fluorine, chlorine, bromine or iodine atom. Further, the term "haloalkyl" is intended to include both monohaloalkyl and polyhaloalkyl. For example, the term "halo(C ₁ -C ₄ )alkyl" is intended to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like. Wait. Unless otherwise specified, examples of haloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl.

The above-described alkyl groups having the specified number of carbon atoms, "alkoxy" represents attached through an oxygen bridge, unless otherwise specified, C _1-6 alkoxy groups include _{C 1, C 2, C 3} , C 4, C 5 , and C ₆ alkoxy groups. In some embodiments, the alkoxy group is a C _1-3 alkoxy group. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentyloxy, and S- Pentyloxy.

Unless otherwise specified, the terms "aromatic ring" and "aryl" are used interchangeably and the term "aryl ring" or "aryl" means a polyunsaturated carbocyclic ring system which may be monocyclic, bicyclic or poly A ring system in which at least one ring is aromatic, and each ring in the bicyclic and polycyclic ring system is fused together. It may be mono- or poly-substituted, may be monovalent, divalent or multivalent, in some embodiments, the aryl is a C _6-12 aryl; in other embodiments, the aryl It is a C _6-10 aryl group. Examples of aryl groups include, but are not limited to, phenyl, naphthyl (including 1-naphthyl and 2-naphthyl, and the like). Substituents for any of the above aryl ring systems are selected from the group of acceptable substituents described herein.

Unless otherwise specified, the terms "heteroaryl ring" and "heteroaryl" are used interchangeably and the term "heteroaryl" means 1, 2, 3 or 4 independently selected from B, N, O and An aryl (or aromatic ring) of a hetero atom of S, which may be a monocyclic, bicyclic or tricyclic ring system in which the nitrogen atom may be substituted or unsubstituted (ie, N or NR, wherein R is H or has been herein Other substituents are defined, and are optionally quaternized, and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O) _p , p is 1 or 2). A heteroaryl group can be attached to the remainder of the molecule through a heteroatom. In some embodiments, the heteroaryl is a 5-10 membered heteroaryl; in other embodiments, the heteroaryl is a 5-6 membered heteroaryl. Examples of the heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl, and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.) , imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.) , triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1,2,4- Triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazole And 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc., pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), benzothiazolyl (including 5-benzothiazolyl, etc.), fluorenyl, Benzimidazolyl (including 2-benzimidazolyl, etc.), fluorenyl (including 5-indenyl, etc.), isoquinolyl (including 1-isoquinolinyl and 5-isoquinolinyl, etc.), Quinoxalinyl (including 2-quinoxalinyl and 5-quinoxalinyl, etc.), Quinolinyl (including 3-quinolyl and 6-quinolinyl, etc.), pyrazinyl, fluorenyl, phenyloxazolyl. Substituents for any of the above heteroaryl ring systems are selected from the group of acceptable substituents described herein.

Unless otherwise specified, C _n-n+m or C _n -C _n+m includes any one of n to n+m carbons, for example, C _1-12 includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ , and C ₁₂ , also including any range of n to n+m, for example, C _1-12 includes C _1-3 , C _1-6 , C _1-9 , C _3-6 , C _3-9 , C _3-12 , C _6-9 , C _6-12 , and C _{9-12 ,} etc.; similarly, n to n The +m element indicates that the number of atoms on the ring is n to n+m, for example, the 3-12 element ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, and a 9-membered ring. a 10-membered ring, an 11-membered ring, and a 12-membered ring, and includes any one of n to n+m, for example, a 3-12-membered ring including a 3-6-membered ring, a 3-9-membered ring, and a 5-6-membered ring. Ring, 5-7 membered ring, 6-7 membered ring, 6-8 membered ring, and 6-10 membered ring.

The term "leaving group" refers to a functional group or atom which may be substituted by another functional group or atom by a substitution reaction (for example, an affinity substitution reaction). For example, representative leaving groups include triflate; chlorine, bromine, iodine; sulfonate groups such as mesylate, tosylate, p-bromobenzenesulfonate, p-toluenesulfonic acid Esters and the like; acyloxy groups such as acetoxy, trifluoroacetoxy and the like.

The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group" or "thiol protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, e.g., tert-butoxycarbonyl (Boc) Arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1, 1-di -(4'-methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS) and the like. The term "hydroxy protecting group" refers to a protecting group suitable for use in preventing hydroxy side reactions. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and t-butyl groups; acyl groups such as alkanoyl groups (e.g., acetyl); arylmethyl groups such as benzyl (Bn), Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and the like.

In the test sample of the present invention, the formate of a compound is obtained by chromatography in a formic acid system (A phase: H2O + 0.225% formic acid, B phase: acetonitrile).

The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, combinations thereof with other chemical synthetic methods, and those well known to those skilled in the art. Equivalent alternatives, preferred embodiments include, but are not limited to, embodiments of the invention.

The solvent used in the present invention is commercially available.

The present invention employs the following abbreviations: DCM stands for dichloromethane; DMF stands for N,N-dimethylformamide; DMSO stands for dimethyl sulfoxide; NMP stands for N-methylpyrrolidone; Boc stands for t-butoxycarbonyl. Amine protecting group; THF stands for tetrahydrofuran; NBS stands for N-bromosuccinimide; TEA stands for triethylamine; DIPEA stands for N,N-diisopropylethylamine; NaOH stands for sodium hydroxide; DBU stands for 1 , 8-diazabicycloundec-7-ene; TFE stands for trifluoroethanol; TFA stands for trifluoroacetic acid; HOBt stands for 1-hydroxybenzotriazole; EDCI.HCl stands for 1-ethyl-(3- Dimethylaminopropyl)carbonyldiimide hydrochloride, NCS stands for N-chlorosuccinimide; EDTA-K ₂ stands for dipotassium ethylenediaminetetraacetate; PEG400 stands for polyethylene glycol 400; PO Representative for oral administration; IV for intravenous administration.

软件命名，市售化合物采用供应商目录名称。 Compound by hand or

Software naming, commercially available compounds using the supplier catalog name.

Detailed ways

The invention is described in detail below by the examples, but is not intended to limit the invention. The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, combinations thereof with other chemical synthetic methods, and those well known to those skilled in the art. Equivalent alternatives, preferred embodiments include, but are not limited to, embodiments of the invention. It will be apparent to those skilled in the art that various changes and modifications may be made to the embodiments of the invention.

Option A

When LR ₇ is H and R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are not OH, the reaction is carried out according to Scheme A.

Compound A1 is subjected to a ring closure reaction with a suitable reagent such as triethyl orthoformate, sulfuric acid/formic acid to give compound A2. Compound A2 is reacted with a suitable chlorinating reagent such as phosphorus oxychloride to give compound A3. Compound A3 is reacted with a Boc protected amine under the action of a suitable base such as TEA or DIPEA to give compound A4. Compound A4 undergoes a deprotection reaction under acidic conditions to give compound A5. If there is no NH ₂ in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ in compound A5, compound A5 is reacted with a suitable acylating reagent such as alkenyl chloride in the presence of a suitable base such as TEA. Compound (I) is obtained; if there is NH ₂ in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ in compound A5, compound A5 is in the presence of a suitable base such as TEA and a suitable acylating agent ( The intermediate compound obtained is reacted with a nitro group at a corresponding position to obtain a compound (I) by a reaction such as alkenyl chloride.

Option B

When LR ₇ is H and there is OH in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ (for example, R ₁ is OH), the reaction proceeds according to Scheme B.

Compound A1 is subjected to a ring closure reaction with a suitable reagent such as triethyl orthoformate, sulfuric acid/formic acid to give compound B1, and then compound B1 is treated with pyridine hydrochloride to give demethylated product B2. Compound B2 is reacted with acetic anhydride in the presence of a suitable base such as pyridine to give compound B3. Compound B3 is reacted with a suitable chlorinating reagent such as phosphorus oxychloride to give compound B4 which is then reacted with a Boc protected amine in the presence of a suitable base such as DIPEA to afford compound B5. Compound B5 and compound B7 are obtained after deacetylating compound B5 and removing the Boc protecting group, respectively. If there is no NH ₂ in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ in compound B7, compound B7 is reacted with a suitable acylating reagent such as alkenyl chloride in the presence of a suitable base such as TEA. Compound (I) is obtained; if there is NH ₂ in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ in compound B7, compound B7 is in the presence of a suitable base such as TEA and a suitable acylating reagent ( The intermediate compound obtained is reacted with a nitro group at a corresponding position to obtain a compound (I) by a reaction such as alkenyl chloride.

Option C

When LR ₇ is other than H and R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are not OH, the reaction proceeds according to Scheme C.

Compound A1 is subjected to a ring closure reaction with a suitable reagent such as urea or a plasma liquid [HDBU ⁺ ] [TFE ^- ] and carbon dioxide gas prepared by DBU and TFE to obtain compound C1. Compound C1 is reacted with a suitable chlorinating reagent such as phosphorus oxychloride to give the dichloro product C2 which is then reacted with a Boc-protected amine in the presence of a suitable base such as DIPEA to afford compound C3. Compound C3 is reacted with a nucleophile such as a substituted amino group, an alcohol or potassium cyanide in the presence of a suitable base such as DIPEA or potassium fluoride to provide compound C4. Deprotection of compound C4 affords compound C5. If there is no NH ₂ in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ in compound C5, compound C5 is reacted with a suitable acylating reagent such as alkenyl chloride in the presence of a suitable base such as TEA. Compound (I) is obtained; if there is NH ₂ in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ in compound C5, compound C5 is in the presence of a suitable base such as TEA and a suitable acylating reagent ( The intermediate compound obtained is reacted with a nitro group at a corresponding position to obtain a compound (I) by a reaction such as alkenyl chloride.

Option D

When LR ₇ is other than H and there is OH in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ (for example, R ₁ is OH), the reaction proceeds according to Scheme D.

Compound A1 is subjected to a ring-closing reaction with a suitable reagent such as urea or a plasma liquid [HDBU ⁺ ][TFE ^- ] and carbon dioxide gas prepared by DBU and TFE to obtain compound D1, and then compound D1 is treated with pyridine hydrochloride to obtain a depolarization. Base product D2. Compound D2 is reacted with acetic anhydride in the presence of a suitable base such as pyridine to afford compound D3. Compound D3 is reacted with a suitable chlorinating reagent such as phosphorus oxychloride to give compound D4 which is then reacted with a Boc-protected amine in the presence of a suitable base such as DIPEA to afford compound D5. Compound D5 is reacted with a nucleophile such as a substituted amino group, an alcohol or potassium cyanide in the presence of a suitable base such as DIPEA or potassium fluoride to give compound D6. Decarboxylation of compound D6 and removal of the Boc protecting group afforded compound D7 and compound D8, respectively. If there is no NH ₂ in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ in compound D8, compound D8 is reacted with a suitable acylating reagent such as alkenyl chloride in the presence of a suitable base such as TEA. Compound (I) is obtained; if there is NH ₂ in R ₁ , R ₂ , R ₃ , R ₄ and R ₅ in compound D8, compound D8 is in the presence of a suitable base such as TEA and a suitable acylating reagent ( The intermediate compound obtained is reacted with a nitro group at a corresponding position to obtain a compound (I) by a reaction such as alkenyl chloride.

Option E

In the above Scheme E, compound E1 is reacted with a suitable acid chloride such as methyl 3-chloro-3-oxopropionate to give compound E2. There are two methods for the synthesis of compound E4: 1 compound E2 and a suitable enamine (such as (E)-4-ethoxy-1,1,1-trifluoro-3-buten-2-one) are condensed to give a compound. E3. The compound E3 is heated and reacted under a dehydrating agent such as p-toluenesulfonic acid to obtain a compound A4 after dehydration, and the compound E2 is directly closed in the presence of a strong base (such as sodium methoxide) to obtain a compound E4. Compound E4 is hydrolyzed to give compound E5, which is then subjected to a Curtius rearrangement reaction to give Boc protected amino compound E6. Compound E6 is deprotected to give compound E7, which is then brominated with the appropriate bromination reagent (e.g., NBS) to afford compound E8. Compound E8 is reacted with a suitable cyanating reagent such as cyanide to give compound E9. Compound E9 is hydrolyzed to give compound A1.

Example 1

first step:

Compound 1a (4.8 g, 28.34 mmol) was dissolved in ethyl acetate (10 mL). After the reaction solution was replaced with hydrogen several times, the reaction was stirred at 15 ° C for 6 hours under a hydrogen balloon. The reaction mixture was filtered, and the filtrate was concentrated to give Compound 1b. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.71-6.62 (m, 3H), 3.88 (s, 3H), 3.76 (brs, 2H).

The second step:

Compound 1b (4.00 g, 28.34 mmol) and TEA (5.74 g, 56.68 mmol) were dissolved in DCM (50 ml), and then 3-chloro-3-oxopropionate was added dropwise at 15 °C ( 5.00 g, 36.62 mmol). After the completion of the dropwise addition, the reaction mixture was further stirred at 15 ° C for 5 minutes and then diluted with DCM (50 mL). The reaction mixture was washed with EtOAc EtOAc EtOAc. The filtrate was concentrated to give the compound 1c. LCMS (ESI) m/z: 264.0 (M+21.

third step:

Compound 1c (6.50 g, 26.95 mmol), (E)-4-ethoxy-1,1,1-trifluoro-3-buten-2-one (4.53 g, 26.95 mmol) and DBU ( 4.31 g, 28.30 mmol) was dissolved in THF (100 mL). The residue was dissolved in EtOAc (EtOAc) (EtOAc)EtOAc. The organic phase was separated, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give the compound 1d.

the fourth step:

Compound 1d (10.50 g, 27.54 mmol) and p-toluenesulfonic acid monohydrate (314.32 mg, 1.65 mmol) were dissolved in toluene (150 mL) and heated to reflux. The water produced by the reaction is separated using a water separator. After the reaction mixture was refluxed for 1 hour, the mixture was refluxed, cooled to 15 ° C, and then washed with water (50 ml), saturated sodium bicarbonate (50 ml) and water (50 ml). The organic phase was collected, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give the compound 1e. LCMS (ESI) m /z:

the fifth step:

Compound 1e (8.20 g, 23.75 mmol) was dissolved in THF (EtOAc) (EtOAc) After the reaction mixture was stirred at 15 ° C for 0.5 hour, a portion of the solvent was removed by pressure and then diluted with water (50 ml). The resulting mixture was washed with methyl tert-butyl ether (80 mL * 2) and the aqueous phase was separated. The aqueous phase was adjusted to pH 2 with concentrated hydrochloric acid and then extracted with ethyl acetate (100 mL*2). The combined organic phases were washed with brine (120 ml), dried over anhydrous sodium sulfate _{^{1 H NMR (400MHz, CDCl 3}} ) δ8.61 (d, J = 7.2Hz, 1H), 7.49-7.43 (m, 1H), 7.05 (d, J = 7.2Hz, 1H), 6.86-6.81 (m, 2H), 3.76 (s, 3H); LCMS (ESI) m.

The sixth step:

Compound 1f (6.30 g, 19.02 mmol) and TEA (2.89 g, 28.53 mmol) were dissolved in tert-butanol (100.00 mL) and then diphenyl azide (6.28 g, 22.82 mmol). The reaction solution was heated to 75 ° C for 2 hours, and then concentrated under reduced pressure. The residue was purified through silica gel column chromatography (EtOAcEtOAcEtOAc LCMS (ESI) m.

Step 7:

The compound 1 g (4.70 g, 11.68 mmol) was dissolved in hydrochloric acid / methanol (50 ml, 4M), and the mixture was stirred at 12 ° C for 13 hours, then concentrated under reduced pressure. The residue was neutralized with aq. EtOAc (EtOAc) The combined extracts were washed with brine (EtOAc) The filtrate was concentrated to give compound 1h. LCMS (ESI) m.

The eighth step:

NBS (64.78 mg, 363.97 μmol) was added to a solution of the compound 1H (100 mg, 330. The mixture was extracted with EtOAc (EtOAc (EtOAc)EtOAc. The filtrate was concentrated under reduced pressure and purified using EtOAc (EtOAc:EtOAc:EtOAc _{^{1 H NMR (400MHz, CDCl 3}} ) δ7.38-7.34 (m, 1H), 6.86 (s, 1H), 6.79-6.73 (m, 2H), 4.92 (brs, 2H), 3.72 (s, 3H); LCMS (ESI) m/z: 38:21.

The ninth step:

Compound 1i (2.60 g, 6.82 mmol) and cuprous cyanide (733.17 mg, 8.18 mmol) were dissolved in NMP (15 mL) and heated to 190 ° C for 4.5 hours in a microwave reactor. The reaction solution was cooled to 20 ° C, then ethyl acetate (30 ml), water (30 ml) and concentrated aqueous ammonia (10 ml). The organic phase was separated, washed with brine (50 ml) The residue was purified by silica gel column chromatography (EtOAc:EtOAc LCMS (ESI) m.

Step 10:

Concentrated sulfuric acid (36.80 g, 375.22 mmol) was diluted with water (5 mL) then compound 1j (1.20 g, 3.67 mmol). The reaction mixture was heated to 80 ° C, stirred for 1 hour, cooled, poured into ice water (200 g), and then adjusted to pH 8 with concentrated aqueous ammonia. The mixture was extracted with EtOAc (EtOAc (EtOAc)EtOAc. The residue was purified by silica gel column chromatography (EtOAcEtOAc LCMS (ESI) m.

The eleventh step:

Compound 1k (0.9 g, 2.61 mmol) was added to triethyl orthoformate (30 ml), and the mixture was heated to 80 ° C for 2 hr. The residue was dissolved in EtOAc (EtOAc)EtOAc. The filtrate was concentrated to give the compound 1l. LCMS (ESI) m/z: 35:21.

Step 12:

Compound 11 (0.9 g, 2.53 mmol) was added to phosphorus oxychloride (10 ml, 107.61 mmol), and the mixture was heated to 80 ° C for 1 hour, and then concentrated under reduced pressure. To the residue was added toluene (15 ml). LCMS (ESI) m.

Step 13:

Compound 1m (1.00 g, 2.68 mmol), tert-butyl piperazine-1-carboxylate (498.41 mg, 2.68 mmol) and TEA (812.36 mg, 8.03 mmol) were dissolved in DCM (20 mL) The reaction was carried out at 15 ° C for 2 hours. Additional TEA (812.36 mg, 8.03 mmol) was added and the reaction was continued at 15 ° C for 16 hours. The reaction mixture was diluted with EtOAc EtOAc EtOAc. The residue was purified by silica gel column chromatography (EtOAcEtOAcEtOAc _{^{1 H NMR (400MHz, CDCl 3}} ) δ8.81 (s, 1H), 7.39-7.34 (m, 1H), 6.89 (s, 1H), 6.80-6.75 (m, 2H), 3.70-3.68 (m, 7H ), 3.59-3.52 (m, 4H), 1.43 (s, 9H); LCMS (ESI) m.

Step 14:

Compound 1n (0.45 g, 859.63 μmol) was added to a hydrochloric acid/methanol solution (20 ml, 4 moles per liter), reacted at 15 ° C for 2 hours, and concentrated to give compound 1o. LCMS (ESI) m/z: 4221.

Step 15:

1o (50 mg, 108.74 micromoles) and TEA (33.01 mg, 326.21 micromoles) were added to DCM (5 mL). After cooling to -30 °C, acryloyl chloride (11.81 mg, 130.48 micromoles) was added. The reaction solution was stirred at -30 ° C for 0.5 hour, then quenched with dilute hydrochloric acid (5 mL, 0.5 moles per liter). The organic phase was separated, dried over anhydrous sodium sulfate, filtered and evaporated. The residue was purified with EtOAc (MeOH:MeOH:EtOAc) The crude product was purified by preparative HPLC (carboxylic acid) to afford Example 1. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.79 (s, 1H), 7.61 - 7.55 (m, 1H), 7.29 (s, 1H), 7.06 (d, J = 8.8 Hz, 1H), 6.96 (t , J = 8.8 Hz, 1H), 6.87-6.80 (m, 1H), 6.30 (dd, J = 16.81, 1.88 Hz, 1H), 5.83 (dd, J = 10.67, 1.88 Hz, 1H), 4.01 (brs, 4H), 3.91 (brs, 4H), 3.84 (s, 3H); LCMS (ESI) m/z: 478.0 (M+1).

Embodiment 2 and Embodiment 3

first step:

Compound 1j (18 g, 55.01 mmol) was dissolved in ionic liquid [HDBU ⁺ ][TFE ^- ] (30.44 g) at 25 ° C, and the reaction was carried out under a balloon filled with carbon dioxide gas for 12 hours, and then the reaction liquid was poured. Into water (100 ml), extracted with ethyl acetate (100 ml *3). The combined organic layers were dried with anhydrous sodium The crude product was added to a mixed solvent of petroleum ether and ethyl acetate (petrole ether: ethyl acetate = 10:1; 15 ml). The filter cake was dried to give compound 2a. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.03-11.34 (m, 2H), 7.67-7.55 (m, 1H), 7.19 (s, 1H), 7.17-7.04 (m, 2H), 3.79 (s) , 3H).

The second step:

Compound 2a (11.4 g, 30.71 mmol) and pyridine hydrochloride (35.49 g, 307.08 mmol) were mixed and heated to 180 ° C, and reacted for 15 minutes and then cooled. The reaction mixture was poured into EtOAc EtOAc m. The organic layers were combined, dried over anhydrous sodium sulfate and filtered, LCMS (ESI) m/z: 35:21.

third step:

Compound 2b (10 g, 27.99 mmol) was dissolved in acetic anhydride (109 g, 100 mL) then pyridine (2.21 g, 27.99 mmol). The reaction mixture was reacted for 10 minutes at 20 ° C, poured into water (50 ml) The combined extracts were dried over anhydrous sodium The residue was added to a mixed solvent of petroleum ether and ethyl acetate ( petroleum ether: ethyl acetate = 8:1; 25 ml).

the fourth step:

Compound 2c (1 g, 2.5 mmol) was dissolved in phosphorus oxychloride (3.84 g, 2.33 ml) and heated to 120 ° C for 0.5 h. The reaction solution was concentrated under reduced pressure to give Compound 2d.

the fifth step:

The synthesis of compound 2e is based on compound 1n. ^{1 H NMR (400MHz, DMSO-} d 6) δ7.70 (dt, J = 8.4,6.4Hz, 1H), 7.50-7.33 (m, 2H), 7.23 (s, 1H), 3.88 (d, J = 3.2 Hz, 4H), 3.57 (s, 4H), 2.11 (s, 3H), 1.45 (s, 9H).

The sixth step:

Compound 2e (150 mg, 0.256 mmol) and TEA (26 mg, 0.256 mmol) were dissolved in methanol (15 mL) and palladium carbon (2.76 mg, 10%). The reaction was allowed to react at 30 ° C for 1 hour under a balloon filled with hydrogen. The reaction solution was filtered and concentrated under reduced pressure. The residue was purified by preparative EtOAc (EtOAc:EtOAc:EtOAc LCMS (ESI) m/z:21.

Step 7:

The synthesis of compound 2g is based on reference compound 1o. LCMS (ESI) m/z: 41:21.

The eighth step:

Compound 2g (80 mg, 0.195 mmol) and TEA (39.55 mg, 0.390 mmol) were dissolved in DCM (10 mL), then acryloyl chloride (17.69 mg, 0.195 mmol) was added dropwise to the reaction at 0 °C. In the liquid. After the completion of the dropwise addition, the reaction was carried out at 20 ° C for 10 minutes, then the reaction mixture was poured into water (20 ml), extracted with ethyl acetate (20 ml * 2), and the organic phase was combined and dried over anhydrous sodium sulfate Concentrated by pressure. The residue was purified by preparative TLC (EtOAc (EtOAc:EtOAc: EtOAc): EtOAc (EtOAc: EtOAc: EtOAc containing 0.05% diethylamine); mobile phase B: carbon dioxide; flow rate: 3mL / min; wavelength: 220nm to give after purification) Example 2 (t _R = 1.763min) and Example 3 (t _R = 1.954min embodiment). LCMS (ESI) m/z: 4621.

Example 2: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.79 (s, 1H), 7.47-7.34 (m, 1H), 7.27 (s, 1H), 6.94-6.71 (m, 3H), 6.29 ( Dd, J = 16.8, 1.6 Hz, 1H), 5.82 (dd, J = 10.4, 1.6 Hz, 1H), 4.09 - 3.87 (m, 8H); LCMS (ESI) m/z: 464.1 (M+1).

Example 3: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.79 (s, 1H), 7.48-7.33 (m, 1H), 7.27 (s, 1H), 6.94-6.72 (m, 3H), 6.29 ( d, J = 16.8 Hz, 1H), 5.83 (d, J = 10.4 Hz, 1H), 4.13 - 3.88 (m, 8H); LCMS (ESI) m/z: 464.1 (M+1).

Example 4

first step:

4-Amino-1-methylpiperidine (38.98 mg, 341.34 μmol) was dissolved in DCM (3 mL). To this solution was added TEA (51.81 mg, 512 μmol) and compound 2e (100 mg, 170.67). Micromolar), then stirred at 15 ° C for 14 hours. A saturated aqueous solution of ammonium chloride (20 ml) was added to the mixture and the mixture was evaporated. The organic phase was dried over anhydrous sodium sulfate and concentrated to give a crude material. This product was purified by preparative TLC (dichloromethane:methanol = 10:1) to afford compound 4a. _{^{1 H NMR (400MHz, CDCl 3}} ) δ7.28-7.19 (m, 1H), 7.06 (br s, 1H), 6.92 (s, 1H), 6.66-6.62 (m, 1H), 4.22-4.11 (m, 1H), 3.62-3.59 (m, 9H), 2.70 (s, 3H), 2.05-2.00 (m, 4H), 1.48 (s, 9H), 1.48-1.38 (m, 4H); LCMS (ESI) m/ z: 621.9 (M + 1).

The second step:

The synthesis of compound 4b is based on compound 1o. LCMS (ESI) m/z: 5221.

third step:

The formate salt of Example 4 was synthesized by reference to Example 1. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.54 (s, 1H), 7.40-7.36 (m, 1H), 7.16 (s, 1H), 6.86-6.77 (m, 3H), 6.28 (dd, J = 2.0, 2.0 Hz, 1H), 5.82 (dd, J=2.0, 1.6 Hz, 1H), 3.97-3.87 (m, 9H), 3.21-3.18 (m, 2H), 2.92 (s, 2H), 2.71 (s) , 3H), 2.22 (d, J = 11.6 Hz, 2H), 1.84-1.81 (m, 2H), 2.49 (s, 3H); LCMS (ESI) m/z: 576.1 (M+1).

Example 5

The formate salt of Example 5 was synthesized by reference to Example 4. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.56 (s, 1H), 7.40-7.34 (m, 1H), 7.21 (s, 1H), 6.83-6.75 (m, 3H), 6.29 (dd, J = 2.0, 2.0 Hz, 1H), 5.82 (dd, J=2.0, 1.6 Hz, 1H), 3.89 (s, 8H), 3.71-3.69 (m, 2H), 3.37 (s, 2H), 3.29-3.28 (m , 2H), 1.27 (t, J = 5.8 Hz, 6H). LCMS (ESI) m.

Example 6

The synthesis of Example 6 is referred to Example 4. _{^{1 H NMR (400MHz, CD 3}} OD) δ7.39-7.33 (m, 1H), 7.15 (s, 1H), 6.86-6.74 (m, 3H), 6.28 (dd, J = 2.0,2.0Hz, 1H) , 5.81 (dd, J = 2.0, 1.6 Hz, 1H), 3.95 (t, J = 4.0 Hz, 9H), 3.90-3.86 (m, 4H), 3.85-3.81 (m, 4H), 3.77-3.75 (m , 4H); LCMS (ESI) m.

Example 7

The synthesis of Example 7 is referred to Example 4. _{^{1 H NMR (400MHz, CD 3}} OD) δ7.39-7.33 (m, 1H), 7.15 (s, 1H), 6.86-6.76 (m, 3H), 6.29 (d, J = 8.4Hz, 1H), 5.82 (d, J = 6.0 Hz, 1H), 4.43-4.16 (m, 2H), 3.96-3.77 (m, 10H), 2.98-2.90 (m, 1H), 2.14 (s, 3H), 2.06 (s, 2H) ), 1.49 (s, 2H); LCMS (ESI) m.

Example 8

The formate salt of Example 8 was synthesized by reference to Example 4. _{^{1 H NMR (400MHz, CD 3}} OD) δ8.48 (s, 1H), 7.39-7.33 (m, 1H), 7.14 (s, 1H), 6.86-6.74 (m, 3H), 6.28 (dd, J = 2.0, 1.6 Hz, 1H), 5.82 (dd, J = 2.0, 2.0 Hz, 1H), 4.40-4.35 (m, 4H), 4.19-4.15 (m, 4H), 3.92-3.90 (m, 1H), 3.90 - 3.86 (m, 8H), 2.87-2.82 (m, 4H), 1.16 (t,J = 6.8 Hz, 6H); LCMS (ESI) m/z: 590.1 (M+1).

Example 9 and Example 10

Example 8 via SFC (column model: Chiralcel OJ-3, 100 x 4.6 mm ID, 3 μm; mobile phase A: methanol (containing 0.05% diethylamine); mobile phase B: carbon dioxide; flow rate: 3 mL/min; wavelength: 220 nm Purified to give Example 9 (t _R = 2.90 min) and

Example 10 (t _R = 3.11 min). LCMS (ESI) m/z:

Example 9: ¹ H NMR (400 MHz, CD ₃ OD) δ 7.36 (dt, J = 8.4, 6.5 Hz, 1H), 7.13 (s, 1H), 6.89-6.71 (m, 3H), 6.28 (dd, J=16.8, 2.0 Hz, 1H), 5.81 (dd, J=10.6, 2.0 Hz, 1H), 4.59 (br s, 1H), 4.40-4.26 (m, 2H), 4.18-4.04 (m, 2H), 3.92-3.71 (s, 9H), 2.71 (q, J = 7.2 Hz, 4H), 1.10 (t, J = 7.2 Hz, 6H); LCMS (ESI) m/z: 590.1 (M+1).

Example 10: ¹ H NMR (400 MHz, CD ₃ OD) δ 7.36 (dt, J = 8.4, 6.5 Hz, 1H), 7.13 (s, 1H), 6.89-6.71 (m, 3H), 6.28 (dd, J=16.8, 2.0 Hz, 1H), 5.81 (dd, J=10.6, 2.0 Hz, 1H), 4.59 (br s, 1H), 4.40-4.26 (m, 2H), 4.18-4.04 (m, 2H), 3.92-3.71 (s, 9H), 2.71 (q, J = 7.2 Hz, 4H), 1.10 (t, J = 7.2 Hz, 6H); LCMS (ESI) m/z: 590.1 (M+1).

Example 11

first step:

Compound 2e (80 mg, 136.53 μmol) and sodium methoxide (29.50 mg, 546.14 μmol) were dissolved in methanol (3 mL) and then stirred at 20 ° C for 30 min. After concentrating the reaction mixture, a crude product was obtained. This product was purified by preparative TLC (dichloromethane:methanol = 10:1) to afford compound 11a. LCMS (ESI) m/?:

The second step:

The synthesis of compound 11b is based on compound 1o.

third step:

The synthesis of Example 11 is referred to Example 1. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.45 (brs, 1H), 7.29-7.21 (m, 1H), 7.13 (s, 1H), 6.77-6.60 (m, 3H), 6.21-6.14 (m, 1H), 5.73-5.68 (m, 1H), 3.96 (s, 3H), 3.92-3.83 (m, 4H), 3.83-3.75 (m, 4H); LCMS (ESI) m/z: 494.0 (M+1) ).

Example 12

first step:

Compound 2e (100 mg, 170.67 μmol) and 1-methylpiperidin-4-ol (196.56 mg, 1.71 mmol) were dissolved in DMSO (2 mL) and dioxane (2 mL) Potassium fluoride (99.16 mg, 1.71 mmol) was added to the solution, and the mixture was heated to 120 ° C and stirred for 2 hours. Water (10 ml) was added to the reaction mixture to quench the mixture, and ethyl acetate (20 ml*2). The organic layer was combined, washed with brine (EtOAc) After the filtrate was concentrated, a crude product was obtained. This product was purified by preparative TLC (dichloromethane:methanol = 10:1) to afford compound 12a. LCMS (ESI) m/z: 6221.

The second step:

The synthesis of compound 12b is based on compound 1o. LCMS (ESI) m/z: 52:21.

third step:

The synthesis of Example 12 is referred to Example 1. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.30-7.21 (m, 1H), 7.14 (s, 1H), 6.75-6.61 (m, 3H), 6.18 (dd, J = 16.0, 4.0 Hz, 1H) , 5.74-5.68 (m, 1H), 5.28-5.14 (m, 1H), 3.91-3.75 (m, 8H), 2.95-2.80 (m, 2H), 2.70-2.55 (m, 2H), 2.41 (s, 3H), 2.15-2.00 (m, 2H), 1.97-1.85 (m, 2H); LCMS (ESI) m/z: 577.2 (M+1).

Example 13

first step:

Potassium cyanide (0.2 g, 3.07 mmol) was dissolved in DMSO (4 mL), and 18-crown-6 (338.33 mg, 1.28 mmol) and compound 2e (150 mg, 256 micromoles) were added to the solution. Then, the reaction was stirred at 15 ° C for 15 hours. The reaction was quenched with EtOAc EtOAc (EtOAc) The organic phase was dried over anhydrous sodium sulfate and concentrated to give a crude material. This product was purified by preparative TLC (petrole ether: ethyl acetate = 1:1) to afford compound 13a. LCMS (ESI) m /z:

The second step:

The synthesis of compound 13b is based on compound 1o. LCMS (ESI) m.

third step:

The formate salt of Example 13 was synthesized by reference to Example 1. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.54 (s, 1H), 7.42 - 7.38 (m, 1H), 7.30 (s, 1H), 6.86-6.74 (m, 3H), 6.31 (dd, J = 1.6, 1.6 Hz, 1H), 5.87-5.80 (dd, J=2.0, 2.0 Hz, 1H), 4.08-4.01 (m, 4H), 3.92-3.88 (m, 4H); LCMS (ESI) m/z: 489.0 (M+1).

Example 14

first step:

Compound 13a (55 mg, 102.91 μmol) was dissolved in dioxane solution (2 mL), and then a solution of hydrochloric acid / dioxane (4.13 ml, 4M) was added to the solution, and then the reaction was stirred at 15 ° C. 1 hour, filtered. The filtrate was concentrated to give the compound 14a. LCMS (ESI) m.

The second step:

The formate salt of Example 14 was synthesized by reference to Example 1. _{^{1 H NMR (400MHz, CD 3}} OD) δ8.55 (s, 1H), 7.31-7.27 (m, 1H), 6.84 (dd, J = 10.4,10.4Hz, 1H), 6.70 (d, J = 8.8Hz , 1H), 6.57-6.22 (m, 1H), 6.32 (dd, J = 2.0, 2.0 Hz, 1H), 5.85 (d, J = 12.8 Hz, 1H), 4.30-4.16 (m, 4H), 3.99- 3.91 (m, 4H); LCMS (ESI) m.

Example 15

The synthesis of Example 15 is referred to Example 12. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.91 (s, 1H), 7.59 (s, 1H), 7.43-7.34 (m, 1H), 7.28 (s, 1H), 6.86-6.74 (m, 3H) , 6.33-6.25 (m, 1H), 5.85-5.79 (m, 1H), 3.99-3.94 (m, 4H), 3.91 (s, 3H), 3.90-3.85 (m, 4H); LCMS (ESI) m/ z: 560.1 (M+1).

Example 16

first step:

Compound 11 (7.00 g, 19.70 mmol) and pyridine hydrochloride (22.77 g, 197.05 mmol) were mixed and then stirred at 180 ° C for 15 min. The reaction mixture was poured into EtOAc EtOAc m. The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give compound 16a.

The second step:

Compound 16a (6.00 g, 17.58 mmol) was dissolved in acetic anhydride (35.9 g, 351.68 mmol) then pyridine (1.39 g, 17.58 mmol). The reaction solution was reacted at 20 ° C for 10 minutes, then poured into water (30 ml) and extracted with ethyl acetate (50 ml*2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give compound 16b.

third step:

The synthesis of compound 16c is based on compound 1m.

the fourth step:

The synthesis of compound 16d is based on compound 1n. LCMS (ESI) m/z: 5621.

the fifth step:

Compound 16d (60.0 mg, 106.1 μmol) was dissolved in THF (3 mL) and water (3 mL). To the solution was added lithium hydroxide monohydrate (251.8 mg, 6.0 mmol), then at 25 ° C Stir for 0.2 hours. The reaction mixture was quenched with EtOAc EtOAc (EtOAc) The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give compound 16e. LCMS (ESI) m/z: Compound:

The sixth step:

The synthesis of compound 16f is based on compound 1o. LCMS (ESI) m/z: 4221.

Step 7:

The synthesis of Example 16 is referred to Example 1. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.75 (s, 1H), 7.41-7.36 (m, 1H), 7.20 (s, 1H), 6.84-6.76 (m, 2H), 6.25 (d, J = 6.0 Hz, 2H), 5.69 (t, J = 2.0 Hz, 1H), 4.43 (d, J = 10.0 Hz, 2H), 4.18 - 4.15 (m, 1H), 3.53-3.46 (m, 2H), 2.71 ( s, 3H), 2.10 (d, J = 11.2 Hz, 2H), 1.73-1.64 (m, 2H); LCMS (ESI) m/z: 478.2 (M+1).

Example 17

The synthesis of Example 17 is referred to Example 16. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.68 (d, J = 3.6 Hz, 1H), 7.41-7.29 (m, 2H), 6.88-6.67 (m, 3H), 6.21-6.08 (m, 1H) , 5.80-5.64 (m, 1H), 4.61 (s, 4H), 4.20-4.13 (m, 2H), 4.12-3.94 (m, 4H), 3.87-3.71 (m, 2H), 2.15 (br s, 2H) LCMS (ESI) m/z: 478.1 (M+1).

Example 18

The synthesis of Example 18 is referred to Example 16. _{^{1 H NMR (400MHz, CD 3}} OD) δ8.68 (s, 1H), 7.53 (s, 1H), 7.44-7.34 (m, 1H), 6.87-6.81 (m, 1H), 6.81-6.71 (m, 1H), 6.59-6.16 (m, 2H), 5.86-5.70 (m, 1H), 5.30-5.05 (m, 1H), 4.77-4.54 (m, 3H), 4.50-4.15 (m, 1H), 4.00- 3.59 (m, 3H), 3.51-3.38 (m, 1H);

Example 19

Compound 19a (22.29 mg, 134.59 μmol), HOBt (9.09 mg, 67.30 μmol) and EDCI.HCl (12.90 mg, 67.30 μmol) were dissolved in DMF (5 mL). TEA (6.81 mg, 67.30 μmol) and compound 2g (30 mg, 67.30 μmol) were then stirred at 25 ° C for 2 hours. The reaction was quenched with EtOAc (EtOAc)EtOAc. The combined organic layers were washed with EtOAc EtOAc. The filtrate was concentrated under reduced pressure to give a crude material. The crude product was purified by preparative TLC (dichloromethane:methanol = 10:1) and preparative HPLC (formic acid) to give the formate salt of Example 19. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.80 (s, 1H), 8.44 (s, 1H), 7.44-7.35 (m, 1H), 7.27 (s, 1H), 6.87-6.74 (m, 4H), 4.05-3.96 (m, 4H), 3.96-3.86 (m, 4H), 3.69-3.64 (m, 2H), 2.68 (s, 6H); LCMS (ESI) m/z: 521.1 (M+1).

Example 20

first step:

To a solution of compound 20a (93.21 g, 567.69 mmol, 93.97 ml, 1.5 eq.) and zinc chloride (2.58 g, 18.92 mmol, 886.29 liters, 0.05 eq.) of acetic anhydride (77.27 g, 756.92 mmol, 70.89 ml, 2 equivalents of dimethyl malonate (50 g, 378.46 mmol, 43.48 ml, 1 equivalent) was added dropwise to the mixture, and the mixture was dropped over 0.5 hr. The reaction solution was heated to 140 ° C and stirred for 1 hour. The reaction mixture was concentrated under reduced vacuo. TLC (petroleum ether: ethyl acetate = 10:1) showed a new dot formation. The reaction mixture was concentrated, and the obtained crystallijjjjjjjj _{^{1 H NMR (400MHz, CDCl 3}} ) δ7.45 (d, J = 12.0Hz, 1H), 7.11 (d, J = 12.4Hz, 1H), 6.25 (t, J = 12.4Hz, 1H), 3.82 (s , 2H), 3.84-3.81 (m, 1H), 3.76 (d, J = 4.0 Hz, 6H).

The second step:

To a solution of compound 20b (28.37 g, 141.70 mmol, 1 eq.) and 2-fluoro-6-methoxy-phenylamine (20 g, 141.70 mmol, 1 eq.) in methanol (150 mL) Sulfonic acid (2.70 g, 14.17 mmol, 0.1 equivalent), the mixture was heated to 80 ° C and stirred for 12 hours. LCMS detected the MS of the target product. The reaction mixture was concentrated to give crystals crystals crystals crystals _{^{1 H NMR (400MHz, CDCl 3}} ) δ7.69-7.54 (m, 2H), 6.94-6.81 (m, 2H), 6.74-6.59 (m, 2H), 6.40 (dt, J = 12.4,2.4Hz, 1H ), 3.83 (s, 3H), 3.76 (s, 3H), 3.71 (s, 3H); LCMS (ESI) m/z: 278.0 (M+1).

third step:

The synthesis of compound 20d is based on compound 1f.

the fourth step:

The synthesis of compound 20e refers to compound 1g. _{^{1 H NMR (400MHz, CDCl 3}} ) δ7.97 (d, J = 7.0Hz, 1H), 7.59 (s, 1H), 7.31 (dt, J = 8.4,6.4Hz, 1H), 6.86-6.67 (m, 3H), 6.21 (t, J = 7.2 Hz, 1H), 3.80-3.69 (m, 3H), 1.49-1.36 (m, 9H).

the fifth step:

The synthesis of compound 20f is based on compound 1h.

The sixth step:

The synthesis of compound 20g is based on compound 1i.

Step 7:

The synthesis of compound 20h is based on compound 1j.

The eighth step:

Heating a mixture of the compound 20 h (1.4 g, 5.40 mmol, 1 eq.), formic acid (5.19 g, 108.01 mmol, 20 eq.) and sulfuric acid (1.59 g, 16.20 mmol, 863.60 μl, 3 eq.) to 100 ° C Stir for 0.5 hours. TLC (petroleum ether: ethyl acetate = 1:1) showed a new dot formation. The reaction mixture was poured into water (30 mL) The combined organic phases were dried over anhydrous sodium sulfate and filtered and evaporated.

The ninth step:

The synthesis of compound 20j is based on compound 2b. LCMS (ESI) m/z:21.

Step 10:

The synthesis of compound 20k is based on compound 2c. LCMS (ESI) m/z: 3121.

The eleventh step:

The synthesis of compound 20l is based on compound 1m.

Step 12:

The synthesis of compound 20m is based on compound 1n. LCMS (ESI) m/z: 4421.

Step 13:

The synthesis of compound 20n is based on compound 1o. LCMS (ESI) m/z: 3421.

Step 14:

The synthesis of Example 20 is referred to Example 1. _{^{1 H NMR (400MHz, CD 3}} OD) δ8.73 (s, 1H), 7.45-7.25 (m, 2H), 6.95-6.75 (m, 4H), 6.28 (dd, J = 16.8,2.0Hz, 1H) , 5.82 (dd, J = 10.6, 2.0 Hz, 1H), 3.90 (s, 8H); LCMS (ESI) m/z: 396.1 (M+1).

Example 21

first step:

To a solution of compound 1c (19.5 g, 80.84 mmol, 1 eq.) in MeOH (100 mL), EtOAc (EtOAc (EtOAc: EtOAc) ML) preparation). The reaction mixture was heated to 70 ° C and allowed to react for 16 hours. LCMS showed the starting material was completely reacted and the MS of the desired product was monitored. The reaction mixture was concentrated, EtOAc mjjjjjjjjj The pH of the organic phase was adjusted to 2 with 35% concentrated hydrochloric acid and then extracted with ethyl acetate (200 mL*3). The combined organic layers were washed with EtOAc EtOAc. The obtained residue was stirred at a mixture of petroleum ether: ethyl acetate = 1 : 2 (30 ml) for 16 hr, and filtered, and the filter cake was dried in vacuo to give Compound 21a. LCMS (ESI) m/z:21.

The second step:

The synthesis of compound 21b refers to compound 1g. LCMS (ESI) m/z: 299.

third step:

The synthesis of compound 21c is based on compound 1h. LCMS (ESI) m/z:21.21.

the fourth step:

The synthesis of compound 21d is based on compound 1i. LCMS (ESI) m.

the fifth step:

The synthesis of compound 21e is based on compound 1j. LCMS (ESI) m/z:21.21.

The sixth step:

The synthesis of compound 21f is based on compound 20i. LCMS (ESI) m/z: 3021.

Step 7:

The synthesis of compound 21g is based on compound 2b. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.10 (s, 1H), 7.44-7.22 (m, 1H), 6.96-6.78 (m, 2H), 6.71 (s, 1H), 2.01 (s, 3H) LCMS (ESI) m/z: 288.1 (M+1).

The eighth step:

The synthesis of compound 21h is based on compound 2c. LCMS (ESI) m.

The ninth step:

The synthesis of compound 21i is based on compound 1m. LCMS (ESI) m/z: 344.0 (M+

Step 10:

The synthesis of compound 21j is based on compound 1n. LCMS (ESI) m.

The eleventh step:

The synthesis of compound 21k is based on compound 1o. LCMS (ESI) m/z: 35:21.

Step 12:

Synthesis of Example 21 Reference Example 1. 1H NMR (400 MHz, CD ₃ OD) δ 8.74 - 8.63 (m, 1H), 8.68 (s, 1H), 7.39 (dt, J = 8.4, 6.6 Hz, 1H) , 6.93-6.79 (m, 3H), 6.69 (s, 1H), 6.29 (dd, J = 16.8, 2.0 Hz, 1H), 5.89-5.78 (m, 1H), 3.89 (s, 8H), 2.17 (s , 3H); LCMS (ESI) m.

Example 22

To a solution of Example 2 (20 mg, 43.16 μmol, 1 eq.) and TEA (5 mg, 49.41 μmol, 6.88 μL, 1.14 eq.) in DCM (2 mL) Methylcarbamoyl chloride (5 mg, 46.49 micromoles, 4.27 microliters, 1.08 equivalents). The above reaction solution was stirred at 0 ° C for 0.5 hour. The target product was detected by LCMS. The reaction mixture was concentrated under reduced pressure. _{^{1 H NMR (400MHz, CD 3}} OD) δ8.79 (s, 1H), 7.69-7.60 (m, 1H), 7.48-7.41 (m, 1H), 7.33 (s, 1H), 7.28-7.19 (m, 1H), 6.88-6.78 (m, 1H), 6.30 (dd, J = 16.8, 2.0 Hz, 1H), 5.83 (dd, J = 10.6, 1.9 Hz, 1H), 4.09-3.96 (m, 4H), 3.95 - 3.85 (m, 4H), 2.89 (s, 3H), 2.74 (s, 3H); LCMS (ESI) m.

Example 23

The synthesis of Example 23 is referred to Example 4. ^{1 H NMR (400MHz, DMSO-} d 6) δ7.40-7.28 (m, 1H), 7.01 (s, 1H), 6.90-6.76 (m, 3H), 6.17 (dd, J = 16.8,2.4Hz, 1H ), 5.83-5.66 (m, 1H), 4.95-4.79 (m, 2H), 4.86 (br d, J = 12.0 Hz, 1H), 3.88-3.48 (m, 8H), 3.04-2.91 (m, 4H) , 2.78 (br s, 5H), 1.92 (br d, J = 11.2 Hz, 2H), 1.47 (br d, J = 8.8 Hz, 2H), 1.09 (br t, J = 7.2 Hz, 6H); LCMS ( ESI) m/z: 618.5 (M + 1).

Example 24

The synthesis of Example 24 was referred to Example 4. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.23 (brd, J = 6.8 Hz, 1H), 6.93 (s, 1H), 6.80 (brd, J = 7.9 Hz, 1H), 6.67 (brt, J = 8.3 Hz, 1H), 6.57 (dd, J = 16.8, 10.6 Hz, 1H), 6.41-6.28 (m, 1H), 5.83-5.71 (m, 1H), 3.81-3.65 (m, 9H), 3.52 ( Br s, 2H), 2.96-2.75 (m, 7H), 2.65-2.47 (m, 2H); LCMS (ESI) m/z: 578.4 (M+1).

Example 25

The synthesis of Example 25 is referred to Example 1 and Example 20. _{^{1 H NMR (400MHz, CD 3}} OD) δ8.79 (s, 1H), 7.33-7.20 (m, 2H), 6.91-6.74 (m, 3H), 6.30 (dd, J = 16.8,2.0Hz, 1H) , 5.83 (dd, J = 10.8, 2.0 Hz, 1H), 4.00 (br s, 4H), 3.91 (br s, 4H), 2.09 (s, 3H); LCMS (ESI) m/z: 460.3 (M+ 1).

Example 26

The synthesis of compound 26a is referred to in Example 1. LCMS (ESI) m/z: 5021.

Compound 26a (1.1 g, 2.19 mmol) was dissolved in ethanol (10 ml) and water (5 ml), and iron powder (611.36 g, 10.95 mmol) and ammonium chloride (1.17 g, 21.89) were added to the solution. Millimol), then stirred at 70 ° C for 1 hour. LCMS showed that the target product was monitored. The mixture was filtered over celite, and the filtered cake was washed with water (20 ml*2). Sodium sulfate (50 g) was dried, filtered and concentrated to give a crude material. This product was purified by preparative HPLC (carboxylic acid) to afford Example 26. ^{1 H NMR (400MHz, DMSO-} d 6) δ8.75 (s, 1H), 7.19 (s, 1H), 6.90 (d, J = 8.0Hz, 1H), 6.78 (dd, J = 10.4,16.4Hz, 1H), 6.71 (d, J = 8.0 Hz, 1H), 6.17 (dd, J = 16.8, 2.0 Hz, 1H), 5.72 (dd, J = 10.4, 2.0 Hz, 1H), 3.88-3.86 (m, 4H) ), 3.79 (br d, J = 13.6 Hz, 4H), 1.82 (s, 3H), 1.72 (s, 3H); LCMS (ESI) m/z: 473.3 (M+1).

Example 27 and Example 28

first step:

Compound 27a (500 mg, 8.12 mmol), acetic anhydride (209.92 mg, 2.06 mmol), 18-crown-6 (27.17 mg, 102.81 mmol) and potassium acetate (100.9 mg, 1.03 mmol) were dissolved in chloroform. (10 ml) was stirred at 25 ° C for 15 minutes, then isoamyl nitrite (361.32 mg, 3.08 mmol) was added, and the mixture was stirred at 75 ° C for 18 hours. The title product was obtained by EtOAc (EtOAc:EtOAc:EtOAc: 3) Extraction, the combined organic layer was washed with brine (20 mL······· This product was purified by column chromatography (ethyl acetate:methanol = 1:1 to 20:1). ^{1 H NMR (400MHz, DMSO-} d 6) δ8.82 (s, 1H), 8.45 (s, 1H), 8.37 (d, J = 8.4Hz, 1H), 7.70 (d, J = 8.8Hz, 1H) , 7.26 (s, 1H), 6.91-6.78 (m, 1H), 6.19 (dd, J = 16.8, 2.0 Hz, 1H), 5.80-5.70 (m, 1H), 3.95-3.73 (m, 8H), 2.73 (s, 3H), 2.18 (s, 3H); LCMS (ESI) m.

The second step:

Example 27 (150 mg, 250.46 μmol) was dissolved in methanol (3 ml), and a solution of hydrochloric acid (0.66 ml) dissolved in water (0.66 ml) was added to the solution, followed by stirring at 25 ° C 30 minute. LCMS showed the title product was formed, and the mixture was concentrated to give a crude product which was purified by preparative HPLC (carboxylic acid) to afford Example 28. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.80 (s, 1H), 7.88 (s, 1H), 7.62 (d, J = 8.4 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H) , 7.23 (s, 1H), 6.84 (dd, J = 16.8, 10.4 Hz, 1H), 6.18 (dd, J = 16.8, 2.4 Hz, 1H), 5.75 (dd, J = 10.4, 2.0 Hz, 1H), 3.92 (br s, 4H), 3.87-3.74 (m, 4H), 2.12 (s, 3H); LCMS (ESI) m/z: 526.2 (M+1).

Example 29, Example 30 and Example 31

The synthesis of Example 29 is referred to Example 26. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.78 (s, 1H), 7.28-7.16 (m, 2H), 6.83 (dd, J = 16.8, 10.6 Hz, 1H), 6.66 (d, J = 8.4 Hz) , 1H), 6.49-6.42 (m, 1H), 6.29 (dd, J = 16.8, 1.9 Hz, 1H), 5.82 (dd, J = 10.6, 2.0 Hz, 1H), 4.05-3.95 (m, 4H), 3.94-3.86 (m, 4H); LCMS (ESI) m.

Example 29 was subjected to SFC (column model: Chiralpak AS-350 x 4.6 mm ID, 3 μm; mobile phase A: methanol (containing 0.05% diethylamine); mobile phase B: carbon dioxide; flow rate: 3 mL/min; wavelength: After isolation and purification at 220 nm), Example 30 (t _R = 1.45 min) and Example 31 (t _R = 1.76 min) were obtained.

Example 30: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.78 (s, 1H), 7.28-7.17 (m, 2H), 6.83 (dd, J = 16.7, 10.6 Hz, 1H), 6.66 (d, J=8.4 Hz, 1H), 6.45 (t, J=8.8 Hz, 1H), 6.34-6.26 (m, 1H), 5.87-5.79 (m, 1H), 4.04-3.95 (m, 4H), 3.94-3.85 (m, 4H); LCMS (ESI) m.

Example 31: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.66 (s, 1H), 7.15-7.04 (m, 2H), 6.71 (dd, J = 16.8, 10.6 Hz, 1H), 6.54 (d, J=8.3 Hz, 1H), 6.33 (t, J=8.9 Hz, 1H), 6.22-6.13 (m, 1H), 5.76-5.62 (m, 1H), 3.90-3.83 (m, 4H), 3.82-3.73 (m, 4H); LCMS (ESI) m.

Example 32 and Example 33

The synthesis of compound 32a is referred to Example 1, Example 2 and Example 26. Compound 32a was subjected to SFC (column model: Chiralpak AS-350 x 4.6 mm ID, 3 μm; mobile phase A: methanol (containing 0.05% diethylamine); mobile phase B: carbon dioxide; flow rate: 3 mL/min; wavelength: 220 nm) After separation and purification, Example 32 (t _R = 2.03 min) and Example 33 (t _R = 2.50 min) were obtained.

Example 32: ¹ H NMR (400 MHz, CD ₃ OD) δ 7.13 - 6.98 (m, 2H), 6.70 (dd, J = 16.8, 10.6 Hz, 1H), 6.53 (d, J = 8.3 Hz, 1H) , 6.32 (t, J = 8.7 Hz, 1H), 6.16 (d, J = 16.6 Hz, 1H), 5.70 (d, J = 10.8 Hz, 1H), 4.25 - 4.12 (m, 2H), 4.03-3.88 ( m, 2H), 3.80-3.67 (m, 8H), 3.64-3.56 (m, 1H), 2.53 (q, J = 6.9 Hz, 4H), 0.96 (t, J = 7.1 Hz, 6H);

LCMS (ESI) m/z: 58:21.

Example 33: ¹ H NMR (400 MHz, CD ₃ OD) δ 7.13 - 6.97 (m, 2H), 6.70 (dd, J = 16.8, 10.6 Hz, 1H), 6.53 (d, J = 8.2 Hz, 1H) , 6.32 (t, J = 8.8 Hz, 1H), 6.16 (d, J = 16.8 Hz, 1H), 5.69 (d, J = 10.6 Hz, 1H), 4.25 - 4.12 (m, 2H), 4.03 - 3.90 ( m, 2H), 3.80-3.67 (m, 8H), 3.64-3.56 (m, 1H), 2.53 (q, J = 6.9 Hz, 4H), 0.96 (t, J = 7.0 Hz, 6H); LCMS (ESI) m/z: 589.4 (M + 1).

Example 34, Example 35, and Example 36

The formate salt of Example 34 was synthesized by referring to Example 2 and Example 26. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.50 (br s, 1H), 7.22 (s, 1H), 6.99 (d, J = 8.4 Hz, 1H), 6.87-6.76 (m, 2H), 6.29 ( Dd, J = 16.8, 2.0 Hz, 1H), 5.85-5.77 (m, 1H), 4.45-4.32 (m, 2H), 4.17 (dd, J = 9.6, 5.6 Hz, 2H), 3.97-3.83 (m, 9H), 2.82 (q, J = 7.2 Hz, 4H), 1.93 (s, 3H), 1.86 (s, 3H), 1.16 (t, J = 7.2 Hz, 6H); LCMS (ESI) m/z: 599.2 (M+1).

Example 34 by SFC (column model: Chiralpak AS-350 x 4.6 mm ID, 3 μm; mobile phase A: methanol (containing 0.05% diethylamine); mobile phase B: carbon dioxide; flow rate: 3 mL/min; wavelength: 220 nm Separation and purification gave Example 35 (t _R = 2.41 min) and

Example 36 (t _R = 3.04 min).

Example 35: ¹ H NMR (400 MHz, CD ₃ OD) δ 7.22 (s, 1H), 6.99 (d, J = 8.4 Hz, 1H), 6.88-6.74 (m, 2H), 6.29 (dd, J = 16.8, 2.0 Hz, 1H), 5.82 (dd, J = 10.8, 2.0 Hz, 1H), 4.40-4.23 (m, 2H), 4.11 (br d, J = 9.6 Hz, 2H), 3.96-3.73 (m, 9H), 2.73 (br d, J = 7.2 Hz, 4H), 1.93 (s, 3H), 1.86 (s, 3H), 1.18-1.16 (m, 1H), 1.18-1.08 (m, 6H); LCMS ( ESI) m/z: 590.3 (M + 1).

Example 36: ¹ H NMR (400 MHz, CD ₃ OD) δ 7.22 (s, 1H), 6.99 (d, J = 8.4 Hz, 1H), 6.88-6.74 (m, 2H), 6.29 (dd, J = 16.8, 2.0 Hz, 1H), 5.82 (dd, J = 10.8, 2.0 Hz, 1H), 4.40-4.23 (m, 2H), 4.11 (br d, J = 9.6 Hz, 2H), 3.96-3.73 (m, 9H), 2.73 (br d, J = 7.2 Hz, 4H), 1.93 (s, 3H), 1.86 (s, 3H), 1.18-1.16 (m, 1H), 1.18-1.08 (m, 6H); LCMS ( ESI) m/z: 590.3 (M + 1).

Example 37

To a solution of Example 34 (40 mg, 66.82 <RTI ID=0.0></RTI></RTI><RTIgt;</RTI><RTIgt; Then, potassium acetate (1.97 mg, 20.04 μmol, 0.3 equivalent) and isoamyl nitrite (15.65 mg, 133.63 μmol, 17.99 μl, 2 equivalent) were added to the above reaction solution. The mixture was stirred at 0 ° C for 0.5 hours and then at 25 ° C for 1.4 hours. TLC (dichloromethane:methanol = 12:1) showed that the starting material was completely reacted, and MS MS of the target compound was detected by LCMS. The reaction mixture was quenched with EtOAc EtOAc. The combined organic phases were washed with brine (10 mL EtOAc) The residue obtained was purified by preparative EtOAc (MeOH:MeOH:EtOAc) _{^{1 H NMR (400MHz, CD 3}} OD) δ7.77 (s, 1H), 7.65 (d, J = 8.0Hz, 1H), 7.44 (d, J = 8.8Hz, 1H), 7.26 (s, 1H), 6.84 (dd, J = 16.8, 10.8 Hz, 1H), 6.29 (dd, J = 16.8, 2.0 Hz, 1H), 5.83 (dd, J = 10.8, 2.0 Hz, 1H), 4.63 (br s, 4H), 4.42-4.29 (m, 2H), 4.14 (dd, J = 5.2, 9.6 Hz, 2H), 3.90-3.847 (m, 9H), 2.77 (q, J = 7.2 Hz, 4H), 2.20 (s, 3H) , 1.13 (t, J = 7.2 Hz, 6H); LCMS (ESI) m.

Example 38

The formate salt of Example 38 was obtained by the synthesis of Reference Example 1, Example 2 and Example 26. _{^{1 H NMR (400MHz, CD 3}} OD) δ8.28 (br s, 1H), 7.26-7.11 (m, 2H), 6.82 (dd, J = 16.8,10.6Hz, 1H), 6.66 (d, J = 8.3 Hz, 1H), 6.45 (t, J = 8.9 Hz, 1H), 6.28 (dd, J = 16.7, 1.8 Hz, 1H), 5.82 (dd, J = 10.6, 1.7 Hz, 1H), 4.48-4.34 (m , 2H), 4.21 (br dd, J = 10.2, 4.8 Hz, 2H), 3.86 (br s, 8H), 3.78-3.66 (m, 1H), 2.59 (s, 6H); LCMS (ESI) m/z : 561.4 (M+1).

Example 39

The formate salt of Example 39 was synthesized by reference to Example 1, Example 2 and Example 26. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.43 (br s, 1H), 7.30-7.14 (m, 2H), 6.83 (dd, J = 16.8, 10.6 Hz, 1H), 6.68 (brd, J = 8.4 Hz, 1H), 6.47 (br t, J = 8.9 Hz, 1H), 6.30 (br d, J = 16.9 Hz, 1H), 5.83 (br d, J = 10.7 Hz, 1H), 4.01-3.85 (m , 10H), 3.42 (br d, J = 4.9 Hz, 2H), 3.35 (s, 3H), 2.92 (s, 6H); LCMS (ESI) m/z: 563.1 (M+1).

Example 40

The formate salt of Example 40 was synthesized by reference to Example 2 and Example 26. _{^{1 H NMR (400MHz, CD 3}} OD) δ8.43 (br s, 1H), 7.20 (s, 1H), 7.11 (t, J = 7.8Hz, 1H), 6.83 (dd, J = 16.8,10.6Hz, 1H), 6.73 (d, J = 7.5 Hz, 1H), 6.63 (d, J = 7.3 Hz, 1H), 6.29 (dd, J = 16.8, 1.8 Hz, 1H), 5.87-5.74 (m, 1H), 4.46-4.32 (m, 2H), 4.21 (dd, J = 10.0, 5.6 Hz, 2H), 4.10-3.92 (m, 1H), 3.87 (br s, 8H), 2.93 (q, J = 7.2 Hz, 4H) ), 1.97 (s, 3H), 1.20 (t, J = 7.3 Hz, 6H); LCMS (ESI) m/z: 585.2 (M+1).

Example 41 and Example 42

In Example 32 (102.32 mg, 164.07 [mu] mol, 1 eq., T _R = 2.03min) was dissolved in acetonitrile (15 mL) was added NCS (28.48 mg, 213.29 mol, 1.3 eq.), The resultant reaction The solution was stirred at 70 ° C for 13 hours. The target product was monitored by LCMS. The reaction was quenched with EtOAc (EtOAc m. The obtained crude product was purified by preparative HPLC (carboxylic acid), and the obtained mixture was further purified from the obtained TLC (dichloroethane:methanol = 10:1) to give Example 41 and Example 42.

Example 41: ¹ H NMR (400 MHz, CD ₃ OD) δ 7.26 (dd, J = 8.93, 5.62 Hz, 1H), 7.03 (s, 1H), 6.71 (dd, J = 16.87, 10.64 Hz, 1H) , 6.38 (t, J = 8.99 Hz, 1H), 6.17 (dd, J = 16.81, 1.90 Hz, 1H), 5.63-5.76 (m, 1H), 4.20 (br t, J = 8.01 Hz, 2H), 3.98 (br d, J = 5.50 Hz, 2H), 3.55 - 3.81 (m, 9H), 2.57 (q, J = 7.09 Hz, 4H); 0.98 (t, J = 7.15 Hz, 6H); LCMS (ESI) m /z: 623.4 (M+1).

Example ^{42: 1 H NMR (400MHz,} CD 3 OD) δ7.14 (t, J = 8.56Hz, 1H), 7.04 (s, 1H), 6.71 (dd, J = 16.75,10.64Hz, 1H), 6.53 (dd, J=8.99, 1.53 Hz, 1H), 6.17 (dd, J=16.75, 1.83 Hz, 1H), 5.70 (dd, J=10.64, 1.83 Hz, 1H), 4.18-4.34 (m, 2H), 3.94-4.11 (m, 2H), 2.70 (br s, 4H), 1.04 (brt, J = 7.09 Hz, 6H); LCMS (ESI) m/z: 563.1 (M+1).

Example 43, Example 44 and Example 45

first step:

NCS (1.39 g, 10.41 mmol, 1.2 eq.) was added dropwise to a solution of compound 32a (5.5 g, 8.67 mmol, 1 eq. 15.5 hours. HPLC showed 46.86% of the remaining material and 34.22% of the target product was formed. Further, NCS (694.75 mg, 5.201 mmol, 0.6 equivalent) was added to the reaction mixture, and the obtained mixture was stirred at 80 ° C for 2 hours. HPLC showed 4.11% of the remaining material and 53.36% of the target product was formed. The reaction mixture was quenched with EtOAc (EtOAc)EtOAc. Purification with methanol = 50:1 to 20:1) gave compound 43a.

The second step:

NCS (64.51 mg, 483.11 μmol, 2 eq.) was added dropwise to a solution of compound 43a (200 mg, 241. HPLC showed the remaining material. The mixture was stirred at 80 ° C for further 12 hours. TLC (dichloroethane:methanol = 10:1) showed that the starting material was completely reacted and the desired product was formed. The reaction mixture was quenched with EtOAc (EtOAc)EtOAc. . The obtained crude product was purified by preparative HPLC (carboxylic acid) to afford Example 43. LCMS (ESI) m/z: 6521.

third step:

Example 43 was chirally resolved by SFC (column model: Cellucoat 50 x 4.6 mm ID, 3 um; mobile phase A: ethanol (containing 0.1% aqueous ammonia) mobile phase B: carbon dioxide; flow rate: 3 mL/min; wavelength: 220 nm ) obtained in Example 44 (t _R = 2.155min) and Example 45 (t _R = 2.361min).

Example ^{44: 1 H NMR (400MHz,} CD 3 OD) δ7.41 (br d, J = 7.2Hz, 1H), 7.04 (s, 1H), 6.70 (dd, J = 16.8,10.6Hz, 1H), 6.16 (d, J = 16.4 Hz, 1H), 5.69 (d, J = 10.4 Hz, 1H), 4.17 (d, J = 7.6 Hz, 2H), 3.97 (s, 2H), 3.73 (d, J = 8.8) Hz, 8H), 3.65-3.54 (m, 1H), 2.53 (q, J = 7.2 Hz, 4H), 0.96 (br t, J = 7.2 Hz, 6H); LCMS (ESI) m/z: 657.2 (M +1).

Example ^{45: 1 H NMR (400MHz,} CD 3 OD) δ7.41 (br d, J = 7.2Hz, 1H), 7.04 (s, 1H), 6.70 (dd, J = 16.8,10.6Hz, 1H), 6.16 (d, J = 16.4 Hz, 1H), 5.69 (d, J = 10.8 Hz, 1H), 4.18 (d, J = 7.6 Hz, 2H), 3.97 (s, 2H), 3.73 (d, J = 8.8) Hz, 8H), 3.65-3.54 (m, 1H), 2.53 (q, J = 7.2 Hz, 4H), 0.96 (br t, J = 7.2 Hz, 6H); LCMS (ESI) m/z: 657.2 (M +1).

Example 46

NCS (181.19 mg, 1.36 mmol, 2 eq.) was added dropwise to a solution of <RTI ID=0.0>#</RTI> </RTI> <RTIgt; . LC-MS showed that the starting material remained and the target product was formed. TLC (dichloroethane:methanol = 10:1) showed that the starting material was completely reacted and three new points were formed. The reaction mixture was stirred with EtOAc EtOAc EtOAc. The obtained mixture was purified by preparative TLC (dichloroethane:methanol = 10:1). LCMS (ESI) m/z: 65:21.

Example 47 and Example 48

To a solution of Example 30 (150 mg, 316.04 [mu] mol, 1 eq., T = 1.45min _R) in acetonitrile (8 mL) under nitrogen was added NCS (33.76 mg, 252.83 mol, 0.8 eq.) And the resulting The mixture was stirred at 70 ° C for 1 hour. LC-MS showed that the target product was generated and TLC showed a new spot. The reaction mixture was poured into water (30 ml), EtOAc (EtOAc) Gram), concentrated by filtration. The residue obtained was purified by preparative EtOAc (MeOH:MeOH:EtOAc)

Example 47: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.66 (s, 1H), 7.28 (dd, J = 5.6, 8.9 Hz, 1H), 7.16 (s, 1H), 6.71 (dd, J = 16.8) , 10.6 Hz, 1H), 6.40 (t, J = 9.0 Hz, 1H), 6.17 (dd, J = 16.8, 1.2 Hz, 1H), 5.76-5.64 (m, 1H), 3.92-3.84 (m, 4H) </ RTI> 3.82-3.73 (m, 4H);

Example 48: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.78 (br s, 1H), 7.35-7.20 (m, 2H), 6.83 (br dd, J = 16.8, 11.4 Hz, 1H), 6.66 (br) d, J = 8.2 Hz, 1H), 6.29 (br d, J = 16.9 Hz, 1H), 5.82 (br d, J = 10.3 Hz, 1H), 4.06 - 3.95 (m, 4H), 3.94 - 3.82 (m , 4H); LCMS (ESI) m.

Example 49

Under nitrogen atmosphere, to acetonitrile (5 ml) Example 30 or 31 (100 mg, 210.69 [mu] mol, 1 eq., T _R = 1.45min) was added NCS (28.13 mg, 210.69 [mu] mol, 1 eq.) Was added, The resulting mixture was stirred at 15 ° C for 2 hours. LC-MS showed the starting material was not completely reacted. The mixture was then stirred at 70 ° C for 2 hours. LC-MS showed the product was detected. The reaction mixture was poured into water (30 ml), EtOAc (EtOAc m. After drying, it was concentrated by filtration. The residue obtained was purified by preparative HPLC (carboxylic acid) to afford Example 49. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.80 (s, 1H), 7.56 (brd, J = 7.2 Hz, 1H), 7.30 (s, 1H), 6.83 (br dd, J = 16.6, 10.6 Hz, 1H), 6.30 (br d, J = 16.6 Hz, 1H), 5.83 (br d, J = 10.6 Hz, 1H), 4.06 - 3.95 (m, 4H), 3.95 - 3.83 (m, 4H); LCMS (ESI) m/z: 531.2 (M+1).

Example 50

The compound 2h (800 mg, 1.73 mmol, 1 eq.) was dissolved in EtOAc (30 mL), then NCS (691.59 mg, 5. . The target product was monitored by LCMS. The reaction was quenched with EtOAc (EtOAc (EtOAc)EtOAc. After drying over anhydrous sodium sulfate, it was concentrated by filtration. The obtained crude product was separated by preparative HPLC (carboxylic acid) to give Example 50. ^{1 H NMR (400MHz, DMSO-} d 6) δ11.37 (br s, 1H), 8.90-8.73 (m, 1H), 7.96 (br s, 1H), 7.22 (s, 1H), 6.83 (dd, J = 16.7, 10.5 Hz, 1H), 6.18 (dd, J = 16.8, 2.3 Hz, 1H), 5.85-5.62 (m, 1H), 3.99-3.70 (m, 8H); LCMS (ESI) m/z: 532.2 (M+1).

Example 51 and Example 52

first step:

Compound 51a was synthesized by reference to Example 29. LCMS (ESI) m/z: 477.1

The second step:

NCS (200.12 mg, 1.50 mmol, 2.1 eq.) was added to a solution of compound 51a (340 mg, s. LC-MS and HPLC showed complete conversion of the starting material and detection of the desired product. The reaction was quenched with EtOAc EtOAc (EtOAc m. The obtained crude product was separated by preparative HPLC (carboxylic acid) to give Compound 51b. LCMS (ESI) m/z: 54: 15.

third step:

Compound 51b was chirally resolved by SFC (column model: DAICEL CHIRALPAK AS (250 mm * 30 mm, 10 um; mobile phase A: ethanol (containing 0.1% aqueous ammonia); mobile phase B: carbon dioxide) to give Example 51 (t _R = 1.569min) and Example 52 (t _R = 2.350min).

Example 51: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.68 (s, 1H), 7.44 (d, J = 7.2 Hz, 1H), 7.07 (s, 1H), 6.81-6.58 (m, 1H) , 6.19 (br dd, J = 16.8, 6.4 Hz, 1H), 5.71 (br d, J = 10.6 Hz, 1H), 4.70 - 4.64 (m, 1H), 4.53 - 3.90 (m, 3H), 3.72-3.34 (m, 2H), 3.17-2.95 (m, 1H), 1.33 (br s, 3H); LCMS (ESI) m/z: 545.1 (M+1).

Example 52: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.8 (s, 1H), 7.44 (d, J = 7.2 Hz, 1H), 7.07 (s, 1H), 6.81-6.46 (m, 1H) , 6.19 (br d, J = 16.4 Hz, 1H), 5.71 (dd, J = 10.8, 1.2 Hz, 1H), 4.64 (br s, 1H), 4.51-4.24 (m, 1H), 4.26-3.84 (m , 2H), 3.68-3.36 (m, 2H), 3.17-2.95 (m, 1H), 1.34 (br s, 3H); LCMS (ESI) m/z: 545.1 (M+1).

Example 53 and Example 54

first step:

Compound 53a was synthesized with reference to compound 51b.

The second step:

Compound 53a was chirally resolved by SFC (column model: DAICEL CHIRALPAK AS (250 mm * 30 mm, 10 um; mobile phase A: ethanol (containing 0.1% aqueous ammonia); mobile phase B: carbon dioxide) to give Example 53 (t _R = 1.429min) and Example 52 (t _R = 2.028min).

Example 53: ¹ H NMR (400 MHz, CD ₃ OD) δ 8.79 (s, 1H), 7.56 (brd, J = 7.2 Hz, 1H), 7.19 (s, 1H), 6.97-6.70 (m, 1H) ), 6.31 (br d, J = 16.0 Hz, 1H), 5.83 (br d, J = 10.4 Hz, 1H), 4.75 (br s, 1H), 4.62-4.27 (m, 2H), 4.26 - 3.97 (m , 1H), 3.79-3.48 (m, 2H), 3.30-3.09 (m, 1H), 1.46 (br s, 3H); LCMS (ESI) m/z: 545.1 (M+1).

Example ^{54: 1 H NMR (400MHz,} CD 3 OD) δ8.80 (s, 1H), 7.56 (d, J = 7.2Hz, 1H), 7.20 (s, 1H), 6.93-6.71 (m, 1H) , 6.31 (br dd, J = 6.0, 16.4 Hz, 1H), 5.83 (dd, J = 10.4, 1.7 Hz, 1H), 4.82-4.77 (m, 1H), 4.61-4.24 (m, 2 H), 4.22 -4.02 (m, 1H), 3.83 - 3.48 (m, 2H), 3.30 - 3.12 (m, 1H), 1.45 (brd, J = 5.2 Hz, 3H); LCMS (ESI) m/z: 545.1 (M +1).

Example 55

first step:

Compound 55a (20 g, 138.73 mmol, 57.14 ml, 1 eq.) was dissolved in THF (200 mL). EtOAc (11.10 g, 277.45 mmol, purity: 60%, 2 eq. After stirring at 0 ° C for 30 minutes, methyl iodide (29.54 g, 208.09 mmol, 12.95 ml, 1.5 eq.) was added, and the obtained mixture was allowed to react at 25 ° C for 18 hours. LC-MS showed a small amount of starting material remaining and the desired product was formed. Water (200 ml) was added to the reaction mixture, and ethyl acetate (300 ml *3). The combined organic layers were dried with anhydrous sodium sulfate and evaporated LCMS (ESI) m / z: 159.0 (M + 1); 1 H NMR (400MHz, CDCl 3) δ7.92-7.85 (m, 3H), 7.59-7.54 (m, 1H), 7.50-7.44 (m, 1H), 7.32-7.25 (m, 2H), 4.05 (s, 3H). LCMS (ESI) m/z: 159.0 (m+1).

The second step:

Compound 55b (10 g, 63.21 mmol, 1 eq.) was dissolved in acetic acid (100 ml), and concentrated nitric acid (6.37 g, 101.14 mmol, 4.55 ml, 1.6 eq.) was added dropwise at 0 ° C. After completion, the reaction system was cooled to 0 ° C and stirred for 1 hour. TLC (petroleum ether: ethyl acetate = 5:1) showed that the starting material was completely reacted. The reaction was poured into a saturated aqueous solution of sodium bicarbonate (1 liter) and ethyl acetate (EtOAc (EtOAc) The organic phase was combined and concentrated under reduced pressure. EtOAc m. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.97 (d, J = 9.17 Hz, 1H), 7.85 (d, J = 8.31 Hz, 1H), 7.73-7.67 (m, 1H), 7.65-7.57 (m, 1H), 7.51-7.43 (m, 1H), 7.35 (d, J = 9.17 Hz, 1H), 4.04 (s, 3H).

third step:

Compound 55c (3 g, 14.76 mmol, 1 eq.) was dissolved in a mixture of ethanol (40 ml) and water (20 ml), and ammonium chloride (7.9 g, 147.64 mmol, 10 eq.) and iron powder. (8.25 g, 147.64 mmol, 10 equivalents), stirred at 90 ° C for 2 hours. LCMS showed the reaction was complete and the target product was detected. The reaction system was filtered and concentrated under reduced pressure to give Compound 55d. LCMS (ESI) m/z: 17.

the fourth step:

Compound 55d (2.5 g, 14.43 mmol, 1 eq.) and potassium carbonate (5.98 g, 43.30 mmol, 3 eq.) were dissolved in acetonitrile (50 mL). 2.96 g, 21.65 mmol, 2.31 ml, 1.5 eq.), stirred at 25 ° C for 12 h. A portion of the starting material was obtained by LCMS. EtOAc (2. EtOAc, 2. LCMS showed the reaction was complete and product formation was detected. The reaction was quenched with EtOAc (EtOAc)EtOAc. The concentrated crude product was slurried for 2 hours (ethyl acetate: petroleum ether = 1:1, 12 ml), filtered, and the filter cake was dried under reduced pressure. Compound 55e was obtained. LCMS (ESI) m/z:21.

the fifth step:

Compound 55e (3.8 g, 11.19 mmol, 1 eq.) was dissolved in MeOH (50 mL) EtOAc. , 16.79 mmol, 2.39 ml, 1.5 eq.) and sodium methoxide (907.01 mg, 16.79 mmol, 1.5 eq.), and the reaction was stirred at 90 ° C for 12 hours. LCMS showed the starting material remained and the reaction was stirred at 90 ° C for a further 6 hours. LCMS showed that the starting material remained, supplemented with 4-ethoxy-1,1,1-trifluoro-3-buten-2-one (940.89 mg, 5.60 mmol, 797.36 μl, 0.5 eq.) and sodium methoxide (302.36 mg, 5.60 mmol, 0.5 eq.), the reaction was stirred at 90 ° C for 15 hours. LCMS showed the reaction was complete and product formation was detected. The reaction mixture was concentrated under reduced vacuo. EtOAc (EtOAc) The organic phases were combined and concentrated under reduced pressure to give a crude compound. LCMS (ESI) m/z: 37:21.

The sixth step:

Compound 55f (4.6 g, 12.19 mmol, 1 eq.) was dissolved in a mixture of water (30 ml) and THF (30 ml), and lithium hydroxide monohydrate (1.02 g, 24.38 mmol, 2 eq.). Stir at 25 ° C for 16 hours. LCMS showed the reaction was complete and the target product was detected. The reaction was quenched by the addition of water (100 mL). EtOAc (EtOAc) The combined organic layers were dried with anhydrous sodium sulfate and evaporated LCMS (ESI) m/z: 36: 39

Step 7:

Compound 55 g (4.4 g, 12.11 mmol, 1 eq.) was dissolved in tert-butanol (50 ml), triethylamine (2.45 g, 24.22 mmol, 3.37 ml, 2 eq.) and 4A molecular sieves (4 g) The resulting mixture was stirred at 90 ° C for 1 hour. Thereafter, DPPA (3.50 g, 12.72 mmol, 2.76 ml, 1.05 equivalent) was added, and stirred at 90 ° C for 1 hour. LCMS showed the reaction was complete and the target product was detected. Filtration, the filtrate was concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography ( silica, petroleum ether: ethyl acetate = 10:1) to afford compound 55h. LCMS (ESI) m/z: 379.1 (M+ ^{1 -} 56); ¹ H NMR (400 MHz, CDCl ₃ -d) δ 8.14 (brd, J = 7.70 Hz, 1H), 8.01 (d, J = 9.05) Hz, 1H), 7.79-7.88 (m, 2H), 7.42-7.48 (m, 1H), 7.34-7.40 (m, 2H), 7.21 (d, J = 8.56 Hz, 1H), 6.93 (d, J = 7.95 Hz, 1H), 3.91 (s, 3H), 1.52 (s, 9H).

The eighth step:

Compound 55h (300 mg, 690.60 μmol, 1 eq.) was dissolved in 1,4-dioxane (4 mL). ), followed by stirring at 25 ° C for 12 hours. LCMS showed some of the starting material remained and was stirred at 45 ° C for 2 hours. LCMS showed little starting material remaining and detection of target product formation. The reaction mixture was concentrated under reduced pressure and evaporated and evaporated. The organic phase was washed with aq. LCMS (ESI) m.

The ninth step:

Compound 55i (1.2 g, 3.59 mmol, 1 eq.) was dissolved in DCM (20 mL), bromosuccinimide (638.90 mg, 3.59 mmol, 1 eq. 0.5 hours. TLC (petroleum ether: ethyl acetate = 3:1) showed the reaction was complete and a new spot was formed. The reaction was quenched with EtOAc (EtOAc)EtOAcEtOAc The combined organic layers were concentrated under reduced vacuo. _{^{1 H NMR (400MHz, CDCl 3}} ) δ8.01 (d, J = 9.05Hz, 1H), 7.85 (d, J = 8.19Hz, 1H), 7.49-7.42 (m, 1H), 7.41-7.33 (m, 2H), 7.25 (d, J = 8.44 Hz, 1H), 7.03 (s, 1H), 5.02 (br s, 2H), 3.92 (s, 3H).

Step 10:

Compound 55j (850 mg, 2.06 mmol, 1 eq.) was dissolved in N,N-dimethylacetamide (20 mL), and then added with zinc powder (1.75 g, 26.74 mmol), Pd ₂ (dba) ₃ (376.76 mg, 411.43 micromoles, 0.2 equivalents), 1,1'-bis(diphenylphosphino)ferrocene (456.18 mg, 822.87 micromoles, 0.4 equivalents) and zinc cyanide (966.25 mg, 8.23 mmol, 522.30 μl, 4 eq.), heated to 120 ° C and stirred for 16 hours. LCMS showed the reaction was complete and the target product was detected. The reaction mixture was filtered, EtOAc (EtOAc) The organic phase was concentrated under reduced pressure. EtOAc m. LCMS (ESI) m / z: 360.2 (M + 1); 1 H NMR (400MHz, CDCl 3) δ8.02 (d, J = 9.17Hz, 1H), 7.86 (d, J = 8.07Hz, 1H), 7.51-7.44 (m, 1H), 7.43 - 7.35 (m, 2H), 7.23 (d, J = 8.44 Hz, 1H), 6.85 (s, 1H), 5.78 (br, s, 2H), 3.92 (s, 3H).

The eleventh step:

Compound 55k (880 mg, 2.45 mmol, 1 eq.) was dissolved in EtOAc (EtOAc) (EtOAcjjjjjjjj LCMS showed the reaction was complete and the target product was detected. The reaction solution was poured into ice water (100 ml), filtered, and then filtered and evaporated. The filter cake was beaten (petroleum ether: ethyl acetate = 1:1, 10 ml) for 1 hour, filtered, and the filter cake was dried under reduced pressure to give compound 55l. LCMS (ESI) m/z: 388.1 (M + 1); ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.06 (br s, 1H), 8.36 (s, 1H), 8.17 (d, J = 9.17 Hz, 1H), 7.99 (d, J = 7.83 Hz, 1H), 7.65 (d, J = 9.17 Hz, 1H), 7.52-7.35 (m, 3H), 7.32 (s, 1H), 3.87 (s, 3H) ).

Step 12:

Compound 55l (600 mg, 1.55 mmol, 1 eq.) was dissolved in phosphorus oxychloride (16.50 g, 107.61 mmol, 10 mL, 69.46 eq.) and N,N-dimethylaniline (938.62 mg, 7.75) Millimol, 981.82 microliters, 5 equivalents), the reaction was heated to stirring for 2 hours. TLC (dichloromethane:methanol = 10:1) showed the reaction was completed. The reaction solution was concentrated under reduced pressure to give a crude compound m.

Step 13:

Compound 55 m (700 mg, 1.73 mmol, 1 eq.) was dissolved in 1,4-dioxane (20 mL). EtOAc (2. EtOAc, EtOAc, And N-Boc piperazine (2.57 g, 13.80 mmol, 8 equivalents), then heated to 50 ° C and stirred for 2 hours. LCMS showed the reaction was complete and the target product was detected. The reaction was quenched with EtOAc EtOAc (EtOAcEtOAcEtOAcEtOAc ), compound 55n was obtained. LCMS (ESI) m / z: 556.5 (M + 1); 1 H NMR (400MHz, CDCl 3) δ8.93 (s, 1H), 8.03 (d, J = 9.05Hz, 1H), 7.86 (d, J =7.46Hz,1H),7.44-7.36(m,3H),7.35-7.30(m,1H),7.05(s,1H),3.90(s,3H),3.83-3.78(m,4H),3.69( Dd, J = 3.85, 6.30 Hz, 4H), 1.52 (s, 9H).

Step 14:

Compound 55n (800 mg, 1.44 mmol, 1 eq.) was dissolved in EtOAc (EtOAc)EtOAc. LCMS showed the reaction was complete and the target product was detected. The reaction solution was concentrated under reduced pressure to give the compound, m. LCMS (ESI) m /z:

Step 15:

Compound 55o (800 mg, 1.40 mmol, 1 eq.) of trifluoroacetic acid salt was dissolved in DCM (15 mL) and EtOAc (1.42 g, 14.05 mmol, 1. Acryloyl chloride (254. 30 mg, 2.81 mmol, 229.10 μl, 2 eq.) was stirred at 0 ° C for 0.5 h. LCMS showed the reaction was complete and the target product was detected. The reaction was quenched with EtOAc (EtOAc)EtOAcEtOAc The organic phases were combined and concentrated under reduced pressure. The obtained crude product was beaten (ethyl acetate: petroleum ether = 1:2, 12 ml), filtered, and the filter cake was dried under reduced pressure to give compound 55p. LCMS (ESI) m/z: 5121.

Step 16:

Compound 55p (200 mg, 392.56 micromoles, 1 eq.) was dissolved in DCM (10 mL). EtOAc (EtOAc, EtOAc, The reaction was carried out for 1 hour. LCMS showed approximately 22.82% product formation. The reaction was quenched with EtOAc (EtOAc)EtOAc. :1) Purification and purification by preparative HPLC (0.075% trifluoroacetic acid) afforded the trifluoroacetate salt of Example 55. LCMS (ESI) m/z: 496.2 (M + 1); ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.33 (br s, 1H), 8.82 (s, 1H), 8.01-7.87 (m, 2H) ), 7.44 - 7.2344 (m, 5H), 6.83 (dd, J = 16.69, 10.45 Hz, 1H), 6.18 (dd, J = 16.69, 2.14 Hz, 1H), 5.81-5.70 (m, 1H), 3.99- 3.73 (m, 8H).

Experimental Example 1: Cell Experiment

Purpose:

This experiment was conducted to demonstrate the inhibitory effect of the compounds of the present invention on KRAS G12C mutant NCI-H358 human non-small cell lung cancer cells, KRAS G12C mutant MIA PaCa2 human pancreatic cancer cells, and wild type A375 human malignant melanoma cells.

Main reagents:

Cell line NCI-H358, cell line A375, cell line MIA Paca2, Cell Titer-Glo assay kit, RPMI1640 medium, DMEM cell culture medium, fetal bovine serum, 0.25% trypsin-EDTA digest, DPBS, cell culture grade DMSO, penicillin

Main instruments:

Multi-label microplate reader Envision, cell culture flask, 384 cell culture microplate, Vi-cell XR cell activity analyzer, CO2 incubator, 300 μL 12-channel electric pipette, Echo ultrasonic nano-liquid workstation

experimental method:

40 μl of phosphate buffer solution was added to the peripheral wells of three 384-well microplates, and 40 μl of the cell suspension to be tested was added to each well of each plate (plate 1: NCI-H358 cell suspension, which contained 500 cells). NCI-H358 cells; plate 2: MIA PaCa2 cell suspension containing 300 MIA PaCa2 cells; plate 3: A375 cell suspension containing 300 A375 cells). The three cell plates were then placed in a carbon dioxide incubator for overnight culture. The Echo test compound was subjected to 3-fold serial dilution, and each compound was diluted by 10 concentration gradients (diluted from 50 μM to 0.003 μM) and 100 nl were added to the corresponding wells of the cell plate, respectively. After the addition, A and P lines, 1 To each of the 24 columns, 40 μL of phosphate buffer was added, and then the cell plate was returned to the carbon dioxide incubator for 5 days. 20 μl of Promega CellTiter-Glo reagent per well was added to the cell plate, and the luminescence signal was stabilized by shaking at room temperature for 10 minutes. Readings were performed using a PerkinElmer Envision multi-label analyzer.

Data Analysis: IC ₅₀ results were analyzed by GraphPad Prism 5.0 IDBS software companies.

Experimental results:

Compounds of the invention NCI-H358 (G12C mutation) cells, anti of A375 (wild-type) cells and MIA PaCa2 (G12C mutant) cell proliferation activity IC ₅₀ data is shown in Table 1 and Table 2.

Conclusion: The compounds of the present invention showed high anti-proliferative activity against KRAS G12C mutant cells NCI-H358 and MIA PaCa2, and the anti-proliferative activity of wild-type A375 cells was weak, showing high selectivity.

Table 1

Test compound NCI-H358 IC ₅₀ (μM) A375 IC ₅₀ (μM) Example 1 5.3 17.8 Example 2 5.36 >50 Example 6 14.64 39.85 Formate for Example 8 1.41 39.09 Example 11 23.99 42.21 Example 12 14.18 9.89 Example 15 13.86 >50 Example 18 7.18 2.86

Example 22 5.12 24.41 Example 26 13.30 32.58 Example 27 20.8 50 Example 28 9.93 36.18 Example 31 1.64 33.29 Example 32 0.45 29.26 Example 33 15.82 39.64 Formate for Example 34 1.05 28.30 Example 37 1.05 19.69 Example 41 0.15 12.24 Example 42 0.01 4.57 Example 44 3.67 9.02 Example 45 0.01 6.24 Example 46 1.95 >50 Example 47 1.00 27.88 Example 48 0.18 20.79 Example 49 0.05 7.32 Example 50 5.49 >50 Example 51 2.04 8.82 Example 52 0.06 7.39 Example 53 2.27 8.91 Example 54 0.29 6.66 Trifluoroacetate salt of Example 55 4.26 50

Table 2

Test compound MIA PaCa2 IC ₅₀ (μM) Example 2 6.48 Example 6 12.7 Formate for Example 8 2.31 Example 25 15.27 Example 31 1.25 Example 32 0.37 Example 35 12.04 Example 36 1.11 Example 37 1.44

Example 41 0.16 Example 42 0.02 Example 44 2.97 Example 45 0.01 Example 46 1.79 Example 47 0.82 Example 48 0.13 Example 49 0.07 Example 50 3.90

Experimental Example 2: Liver microsome stability test

Purpose:

The metabolic stability of the test article in mouse, rat and human liver microsomes was tested.

Experimental Materials:

Test article (10 mM), Testosterone (testosterone, control, 10 mM), Diclofenac (diclofenac, control, 10 mM), Propafenone (propylpropionide, control, 10 mM), human liver microsomes, rat liver microsomes, Mouse liver microsomes.

Buffer system:

1. 100 mM potassium phosphate buffer (pH 7.4).

2. 10 mM magnesium dichloride solution.

Compound dilution:

1. Intermediate solution: 5 μL of the test sample or control substance was diluted with 45 μL of DMSO (with 450 μL of 1:1 methanol/water).

2. Working solution: The intermediate solution was diluted with 450 μL of 100 mM potassium phosphate buffer.

NADPH regeneration system:

1. β-phosphoamide adenine dinucleotide, derived from Sigma, Cat. No. N0505.

2. Isocitric acid, from Sigma, catalog number I1252.

3. Isocitrate dehydrogenase, from Sigma, Cat. No. I2002.

Preparation of liver microsome solution (final concentration: 0.5 mg protein/mL):

Stop solution:

Cold acetonitrile containing 100 ng/mL Tolbutamide and 100 ng/mL Labetalol (Laberolol) was used as an internal standard.

experimental method:

1. Add 10 μL of the test or control working solution to all plates (T0, T5, T10, T20, T30, T60, NCF60).

2. Dispense 680 μL/well of liver microsome solution into a 96-well plate, then add 80 μL/well to each plate, and place the above incubator plate at 37 ° C for about 10 minutes.

3. Add 10 μL of 100 mM potassium phosphate buffer to each well on the NCF60 plate.

4. After the pre-incubation, dispense 90 μL/well of NADPH regeneration system working solution to a 96-well plate, then add 10 μL/well l to each plate to initiate the reaction.

5. Incubate for the appropriate time (eg 5, 10, 20, 30 and 60 minutes).

6. Add 300 μL of Stop Solution (refrigeration at 4 ° C with 100 ng/mL Tolbutamide and 100 ng/mL Labetalol) to each sample well.

7. Shake the sample plate for approximately 10 minutes and centrifuge at 4000 rpm for 20 minutes at 4 degrees.

8. When centrifuging, add 300 μL of HPLC water to each well and take 100 μL of the supernatant for LC-MS/MS analysis.

data analysis:

T _1/2 and Cl _int(mic) are calculated by the following formula.

when

Each gram of liver contains 45 mg of microsomal protein, and the liver weights of mice, rats, dogs, monkeys, and humans are 88 g/kg, 40 g/kg, 32 g/kg, 30 g/kg, and 20 g/kg, respectively.

C _t is the concentration at time t, t is the incubation time, the concentration when C ₀ is 0, K _e is the elimination rate constant, Cl _{int (mic)} is the intrinsic clearance of liver particles, and Cl _{int (liver)} is the intrinsic clearance of liver rate.

CL _int(mic) =0.693/half-life/mg microsomal protein per mL (microsomal concentration at incubation)

CL _int(liver) =CL _int(mic) ×mg microsomal protein / g liver weight × liver weight to body weight ratio

Experimental results: See Table 3.

Experimental results:

The compound of the present invention shows a long half-life in the liver microsome stability test of human, rat and mouse, and therefore it is presumed that the compound of the present invention has good metabolic stability in vivo.

table 3

Experimental Example 3: Rat pharmacokinetic evaluation experiment

Purpose:

Male Sprague-Dawley rats were used as test animals, and the concentration of the drug in plasma at different times after intravenous administration and administration of the test compound was determined by LC/MS/MS. The pharmacokinetic behavior of the test compound in rats was studied and its pharmacokinetic characteristics were evaluated.

Experimental protocol: Test animals: 10 healthy adult male Sprague-Dawley rats were divided into 4 groups according to the principle of similar body weight. Group IV (two groups) each group of 2, PO group (two groups) each group of 3. Animals were purchased from Beijing Weitong Lihua Experimental Animal Co., Ltd.

Drug preparation:

Group IV: Weigh the appropriate amount of sample, add appropriate amount of DMSO, PEG400 and water according to the volume ratio of 10:60:30, and stir to obtain a clarified state of 1.5 mg/mL.

PO group: Weigh the appropriate amount of sample, add appropriate amount of DMSO, PEG400 and water according to the volume ratio of 10:60:30, and stir to obtain a clarified state of 1.0 mg/mL.

Dosing:

After fasting overnight, group IV was administered intravenously at a dose of 2 mL/kg at a dose of 3 mg/kg. The PO group was administered intragastrically at a dose of 10 mL/kg and a dose of 10 mg/kg.

Experimental operation:

Male SD rats were given the test compound, and 200 ul of blood was collected at 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours, and placed in commercialization with EDTA-K ₂ in advance. In the anti-coagulation tube. After administration of the test compound to the intragastric administration group, 200 ul of blood was collected at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours, respectively, and placed in a commercial anticoagulant tube to which EDTA-K _{2 was} previously added. in. The tube was centrifuged for 15 minutes to separate the plasma and stored at -60 °C. Animals can be fed after 2 hours of administration. The content of test compounds in plasma after intravenous and intragastric administration in rats was determined by LC/MS/MS. The linear range of the method was 2.00-6000 nM; plasma samples were analyzed by acetonitrile precipitation protein analysis.

Experimental results:

The experimental results are shown in Table 4.

Experimental results:

In the rat pharmacokinetic evaluation experiment, the compounds of the present invention showed higher exposure and better oral availability than the reference compound ARS-1620.

Table 4

Note: Cl: clearance rate; V _d : volume of distribution; AUC: exposure; T _1/2 : half-life; C _max : maximum concentration of compound after oral administration; T _max : time to reach C _max ; F: bioavailability degree.

Experimental Example 4: In vivo efficacy test (1)

Purpose:

The in vivo efficacy of the test compound on the subcutaneous xenograft tumor model of human pancreatic cancer MIA-PaCa2 cells was evaluated.

Experimental operation:

BALB/c nude mice, female, 6-8 weeks, weigh approximately 18-22 grams. Each mouse was inoculated subcutaneously with 0.2 mL (1 × 10 ⁷ ) MIA-PaCa 2 cells (with matrigel, volume ratio 1:1). Dosing begins when the average tumor volume reaches approximately 169 cubic millimeters. The test compound was orally administered daily as shown in Table 5. The tumor volume was measured twice a week and the volume was measured in cubic millimeters and was calculated by the following formula: V = 0.5 a x b ² , where a and b are the long and short diameters of the tumor, respectively. The antitumor effect of the compound was evaluated by TGI (%). TGI (%), reflecting the tumor growth inhibition rate. Calculation of TGI (%): TGI (%) = [(1 - mean tumor volume at the end of administration of a treatment group - mean tumor volume at the start of administration of the treatment group)) / (average tumor at the end of treatment of the solvent control group) Volume-solvent control group average tumor volume at the start of treatment)] × 100%. Experimental results: See Table 5.

table 5

Group Tumor volume (mm ³ ) (Day 20) TGI (%) Solvent control 612±75 -- Example 2 (50 mg/kg) 457±94 35 Example 2 (200 mg/kg) 307±61 69

Experimental results:

The compounds of the invention exhibit good in vivo efficacy in a human pancreatic cancer MIA-PaCa2 cell subcutaneous xenograft model. Twenty days after the start of administration, the compound of the present invention had a significant antitumor effect as compared with the solvent control group, and had a significant dose-effect relationship.

Experimental Example 5: In vivo efficacy test (2)

Purpose:

The in vivo efficacy of the test compound on the human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model was evaluated.

Experimental operation:

BALB/c nude mice, female, 6-8 weeks old, weighing 18-21 grams. A total of 100 are required. Provided by Shanghai Lingchang Experimental Animal Co., Ltd. The NCI-H358 tumor cells were resuspended in PBS to prepare 0.1 mL (5×10 ⁶ ) cell suspensions, which were subcutaneously inoculated into the right back of each mouse (5×10 ⁶ /piece) for tumor growth. . Randomized dosing was initiated when the average tumor volume reached approximately 150-200 mm ³ as shown in Table 6. Tumor diameters were measured twice a week using vernier calipers. The calculation formula of tumor volume is: V = 0.5a × b ² , and a and b represent the long diameter and short diameter of the tumor, respectively. The antitumor effect of the compound was evaluated by TGI (%). TGI (%), reflecting the tumor growth inhibition rate. Calculation of TGI (%): TGI (%) = [(1 - mean tumor volume at the end of administration of a treatment group - mean tumor volume at the start of administration of the treatment group) / (mean tumor volume at the end of treatment of the solvent control group) - The average tumor volume at the start of treatment in the solvent control group)] x 100%.

Experimental results: See Table 6.

Table 6

Experimental Conclusion: The compounds of the present invention exhibited good in vivo efficacy in a human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model. Twenty days after the start of administration, the compound of the present invention has a significant antitumor effect as compared with the reference compound ARS-1620.

Experimental Example 6: In vivo efficacy test (3)

Purpose:

The in vivo efficacy of the test compound on a human pancreatic cancer x-MIA-PaCa2 cell subcutaneous xenograft tumor model was evaluated.

Experimental operation:

NU/NU mice, female, 6-8 weeks old, weighing 17-20 grams. A total of 100 (more than 30% of the animals) are required. Provided by Beijing Weitong Lihua Technology Co., Ltd. 0.2 mL (10×10 ⁶ ) of x-MIA-PaCa 2 cells (with matrigel, volume ratio of 1:1) were subcutaneously inoculated into the right back of each mouse, and the group was started when the average tumor volume reached about 150 mm ³ . The dose was administered as shown in Table 7. Tumor diameters were measured twice a week using vernier calipers. The calculation formula of tumor volume is: V = 0.5a × b ² , and a and b represent the long diameter and short diameter of the tumor, respectively. The antitumor effect of the compound was evaluated by TGI (%). TGI (%), reflecting the tumor growth inhibition rate. Calculation of TGI (%): TGI (%) = [(1 - mean tumor volume at the end of administration of a treatment group - mean tumor volume at the start of administration of the treatment group) / (mean tumor volume at the end of treatment of the solvent control group) - The average tumor volume at the start of treatment in the solvent control group)] x 100%.

Experimental results: See Table 7.

Table 7

Experimental Conclusion: The compounds of the present invention show good in vivo efficacy in a human pancreatic cancer x-MIA-PaCa2 cell subcutaneous xenograft model. The compound of the present invention had a significant antitumor effect compared to the reference compound ARS-1620 14 days after the start of administration.

Experimental Example 7: In vivo efficacy test (4)

Purpose:

The in vivo efficacy of the test compound on the human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model was evaluated.

Experimental operation:

BALB/c nude mice, female, 6-8 weeks old, weighing 18-20 grams. A total of 40 are needed. Provided by Shanghai Lingchang Experimental Animal Co., Ltd. NCI-H358 tumor cells were resuspended in PBS to prepare a cell suspension with a density of 5 × 10 ⁷ /mL, and subcutaneously inoculated into the right back of each mouse (0.1 mL, 5 × 10 ⁶ /pc) Waiting for tumor growth. When the average tumor volume reached about 166 mm ³ , randomized administration was started, and the doses are shown in Table 8. Tumor diameters were measured twice a week using vernier calipers. The calculation formula of tumor volume is: V = 0.5a × b ² , and a and b represent the long diameter and short diameter of the tumor, respectively. The antitumor effect of the compound was evaluated by TGI (%). TGI (%), reflecting the tumor growth inhibition rate. Calculation of TGI (%): TGI (%) = [(1 - mean tumor volume at the end of administration of a treatment group - mean tumor volume at the start of administration of the treatment group) / (mean tumor volume at the end of treatment of the solvent control group) - The average tumor volume at the start of treatment in the solvent control group)] x 100%.

Experimental results: See Table 8.

Table 8

Experimental conclusion: After 27 days from the start of administration, the compound of the present invention has a significant antitumor effect compared to the reference compound ARS-1620 at the same dose (15 mg/kg). In addition, the compound of the present invention exhibited a remarkable tumor suppressing effect at a dose (5 mg/kg) lower than that of the reference compound ARS-1620 (15 mg/kg). This indicates that the compound of the present invention exhibits good in vivo efficacy in a human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model, and the antitumor effect has a dose-dependent tendency.

Claims (26)

a compound of the formula (I), a pharmaceutically acceptable salt thereof or an isomer thereof,

among them, Ring A is selected from 3 to 8 membered heterocycloalkyl, and said 3 to 8 membered heterocycloalkyl is optionally substituted by 1, 2 or 3 R; R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from the group consisting of H, halogen, OH, NH ₂ , CN, C _1-6 alkyl and C _1-6 heteroalkyl, said C _{1- 6} alkyl and C _1-6 heteroalkyl are optionally substituted by 1, 2 or 3 R; Or, R ₁ and R _{2 are} joined together to form a ring B; Or R ₂ and R _{3 are} joined together to form a ring B; Or R ₃ and R _{4 are} joined together to form a ring B; Or R ₄ and R _{5 are} joined together to form a ring B; Ring B is selected from the group consisting of phenyl, C _5-6 cycloalkenyl, 5- to 6-membered heterocycloalkenyl, and 5- to 6-membered heteroaryl, phenyl, C _5-6 cycloalkenyl, and 5 to 6-membered hetero a cycloalkenyl group, a 5- to 6-membered heteroaryl group, optionally substituted by 1, 2 or 3 R _a ; R _{a is} selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-6 alkyl and C _1-6 heteroalkyl, the C _1-6 alkyl and C _1-6 heteroalkyl optionally being 1, 2 Or 3 R substitutions; R ₆ is selected from H, halogen and C _1-6 alkyl, said C _1-6 alkyl optionally substituted with 1, 2 or 3 R &lt; R _{7 is} selected from the group consisting of H, CN, NH ₂ , C _1-8 alkyl, C _1-8 heteroalkyl, 4-6-membered heterocycloalkyl, 5- to 6-membered heteroaryl, and C _5-6 cycloalkyl , the C _1-8 alkyl group, the C _1-8 heteroalkyl group, the 4-6 membered heterocycloalkyl group, the 5- to 6-membered heteroaryl group, and the C _5-6 cycloalkyl group are selected from the group consisting of 1, 2 Or 3 R substitutions; L is selected from the group consisting of a single bond, -NH-, -S-, -O-, -C(=O)-, -C(=S)-, -CH ₂ -, -CH(R _b )-, and -C( R _b ) ₂ -; L' is selected from the group consisting of a single bond and -NH-; R _{b is} selected from C _1-3 alkyl and C _1-3 heteroalkyl, and the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted by 1, 2 or 3 R; R _{8 is} selected from H, C _1-6 alkyl and C _1-6 heteroalkyl, and the C _1-6 alkyl and C _1-6 heteroalkyl are optionally substituted by 1, 2 or 3 R; R is selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-6 alkyl, C _1-6 heteroalkyl, and C _3-6 membered cycloalkyl, said C _1-6 alkyl, C _1-6 hetero An alkyl group and a C _3-6 membered cycloalkyl group are optionally substituted by 1, 2 or 3 R'; R' is selected from the group consisting of: F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , CH ₃ O, CF ₃ , CHF ₂ , CH ₂ F, cyclopropyl, propyl, iso Propyl, N(CH ₃ ) ₂ , NH(CH ₃ ); "Heter" means a hetero atom or a hetero atom group, the 3- to 8-membered heterocycloalkyl group, a C _1-6 heteroalkyl group, a 5- to 6-membered heterocycloalkenyl group, a 5- to 6-membered heteroaryl group, and C _{1 - 8} The "hetero" of a heteroalkyl group, a 4-6 membered heterocycloalkyl group, and a _C1-3 heteroalkyl group are each independently selected from -C(=O)N(R)-, -N(R)-, -NH. -, N, -O-, -S-, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O ₂ - and -N(R)C(=O)N(R)-; In any of the above cases, the number of heteroatoms or heteroatoms is independently selected from 1, 2 and 3. The compound according to claim 1, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R is selected from the group consisting of F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , CH ₃ O, CF ₃ , CHF ₂ , CH ₂ F, cyclopropyl, propyl, isopropyl, N(CH ₃ ) ₂ , NH(CH ₃ ) and N(CH ₂ CH ₃ ) ₂ . The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein ring A is selected from the group consisting of aziridine, azetidine, pyrrolidine, piperidinyl, piperazine a 1,4-diazacycloheptyl group and a 3,6-diazabicyclo[3.2.0]heptane, the aziridine, azetidine, pyrrolidine, piperidinyl, Piperazinyl, 1,4-diazacycloheptyl and 3,6-diazabicyclo[3.2.0]heptane are optionally substituted by 1, 2 or 3 R. The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from H, F, and Cl , Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH and CH ₃ NH(C=O)O, the CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH and CH ₃ NH(C=O)O are optionally substituted by 1, 2 or 3 R. The compound according to claim 4, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from H, F, Cl, Br , I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ N (C=O O and CH ₃ NH(C=O)O. The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein ring B is selected from pyrazolyl, imidazolyl, pyrrolyl, thienyl, furyl, triazolyl, Oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, phenyl, pyridyl, pyrimidinyl, pyridazinyl, triazinyl, morpholinyl, cyclopentenyl and cyclohexenyl, said pyridyl Azyl, imidazolyl, pyrrolyl, thienyl, furyl, triazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, phenyl, pyridyl, pyrimidinyl, pyridazinyl, triazine The base, morpholinyl, cyclopentenyl and cyclohexenyl are optionally substituted by 1, 2 or 3 R _a . The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R _{a is} selected from the group consisting of F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ C(=O). The compound according to claim 6 or 7, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein ring B is selected from the group consisting of phenyl, pyrazolyl, 1-methyl-1H-pyrazolyl and 1- (1H-pyrazol-1-yl)ethanone. The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R _{6 is} selected from the group consisting of H, F, Cl, Br, I and C _1-3 alkyl, said C _{The 1-3} alkyl group is optionally substituted by 1, 2 or 3 R groups. The compound according to claim 9, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R _{6 is} selected from the group consisting of H, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ , CH ₂ F . The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R _{7 is} selected from the group consisting of H, CN, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkane , morpholinyl, piperidinyl, azetidinyl, azetidinyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, cyclohexane , cyclopentyl, phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, said C _1-6 alkyl, C _1-6 heteroalkyl, morpholinyl, piperidinyl, nitrogen Heterocyclobutane, azetidinyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, cyclohexane, cyclopentyl, phenyl, pyridine The base, pyridazinyl, pyrimidinyl, pyrazinyl is optionally substituted by 1, 2 or 3 R. The compound according to claim 11, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R _{7 is} selected from the group consisting of H, CH ₃ , CN, NH ₂ ,

Said

Optionally substituted by 1, 2 or 3 R. The compound according to claim 12, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R _{7 is} selected from the group consisting of H, CH ₃ , CN, NH ₂ ,

The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein R _{8 is} selected from the group consisting of H, C _1-4 alkyl and C _1-4 heteroalkyl, said C _{The 1-4} alkyl group and the C _1-4 heteroalkyl group are optionally substituted by 1, 2 or 3 R groups. The compound according to claim 14, or a pharmaceutically acceptable salt thereof, or an isomer thereof, wherein R _{8 is} selected from the group consisting of H, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CHCH ₂ , (CH ₃ ) ₂ CH, CH ₃ O, CH ₃ NH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ NCH ₂ and CH ₃ NHCH ₂ . The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein the structural unit

Selected from

Wherein R _{9 is} selected from the group consisting of H and C _1-3 alkyl. The compound according to claim 16, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein the structural unit

Selected from

The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein the structural unit

Selected from H, CN, CH ₃ , CH ₃ CH ₂ , (CH ₃ ) ₂ CH, (CH ₃ ) ₂ N, (CH ₃ ) ₂ NCH ₂ ,

The compound according to claim 1 or 2, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein the structural unit

Selected from

The compound according to any one of claims 1 to 19, a pharmaceutically acceptable salt thereof or an isomer thereof, selected from the group consisting of

Where L is as defined in claim 1 R ₁ , R ₂ , R ₄ and R _{5 are} as defined in claim 1 or 4, R _{6 is} as defined in claim 1 or R _{, 7} as defined in claim 1, 11, 12 or 13 R _{, 8} as defined in claim 1, 14 or 15 R _{, 9 is} as defined in claim 16. The compound according to claim 20, a pharmaceutically acceptable salt thereof or an isomer thereof, selected from

Wherein R ₁ , R ₂ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ and L are as defined in claim 20, and ring B is as defined in claim 1 or 6. The compound according to claim 21, a pharmaceutically acceptable salt thereof or an isomer thereof, selected from

Wherein R ₁ , R ₂ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , L, R ₉ and R _{a are} as defined in claim 21. a compound of the formula: a pharmaceutically acceptable salt thereof or an isomer thereof, selected from

The compound according to claim 23, a pharmaceutically acceptable salt thereof or an isomer thereof, selected from

The use of a compound according to any one of claims 1 to 24, a pharmaceutically acceptable salt thereof or an isomer thereof for the preparation of a medicament for treating cancer. The use according to claim 25, wherein the cancer comprises lung cancer, lymphoma, esophageal cancer, ovarian cancer, pancreatic cancer, rectal cancer, glioma, cervical cancer, urothelial carcinoma, gastric cancer, intrauterine Membrane cancer, liver cancer, cholangiocarcinoma, breast cancer, colon cancer, leukemia and melanoma.